U.S. patent application number 15/117982 was filed with the patent office on 2016-12-15 for pyrazolo[1,5-a]pyrimidine-5,7-diamine compounds as cdk inhibitors and their therapeutic use.
The applicant listed for this patent is CANCER RESEARCH TECHNOLOGY LIMITED, EMORY UNIVERSITY, IMPERIAL INNOVATIONS LIMITED. Invention is credited to Simak ALI, Anthony BARRETT, Alexander BONDKE, Charles COOMBES, Matthew FUCHTER, Sebastian KROLL, Brian SLAFER, James Patrick SNYDER.
Application Number | 20160362410 15/117982 |
Document ID | / |
Family ID | 50482596 |
Filed Date | 2016-12-15 |
United States Patent
Application |
20160362410 |
Kind Code |
A1 |
BONDKE; Alexander ; et
al. |
December 15, 2016 |
PYRAZOLO[1,5-A]PYRIMIDINE-5,7-DIAMINE COMPOUNDS AS CDK INHIBITORS
AND THEIR THERAPEUTIC USE
Abstract
The present invention pertains generally to the field of
therapeutic compounds. More specifically the present invention
pertains to certain pyrazolo[1,5-a]pyrimidine-5,7-diamine compounds
(referred to herein as "PPDA compounds") that, inter alia, inhibit
(e.g., selectively inhibit) CDK (e.g., CDK1, CDK2, CDK4, CDK5,
CDK6, CDK7, CDK8, CDK9, CDK10, CDK11, CDK12, CDK13, etc.). The
present invention also pertains to pharmaceutical compositions
comprising such compounds, and the use of such compounds and
compositions, both in vitro and in vivo, to inhibit CDK; and to
treat disorders including: disorders that are associated with CDK;
disorders that result from an inappropriate activity of a
cyclin-dependent kinase (CDK); disorders that are associated with
CDK mutation; disorders that are associated with CDK
overexpression; disorders that are associated with upstream pathway
activation of CDK; disorders that are ameliorated by the inhibition
of CDK; proliferative disorders; cancer; viral infections
(including HIV); neurodegenerative disorders (including Alzheimer's
disease and Parkinson's disease); ischaemia; renal diseases; and
cardiovascular disorders (including atherosclerosis). Optionally,
the treatment further comprises treatment (e.g., simultaneous or
sequential treatment) with a further active agent which is, e.g.,
an aromatase inhibitor, an anti-estrogen, a Her2 blocker, a
cytotoxic chemotherapeutic agent, etc.
Inventors: |
BONDKE; Alexander; (Caputh,
DE) ; KROLL; Sebastian; (London, Greater London,
GB) ; BARRETT; Anthony; (London, Greater London,
GB) ; FUCHTER; Matthew; (London, Greater London,
GB) ; SLAFER; Brian; (Lemont, IL) ; ALI;
Simak; (London, Greater London, GB) ; COOMBES;
Charles; (London, Greater London, GB) ; SNYDER; James
Patrick; (Atlanta, GA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
CANCER RESEARCH TECHNOLOGY LIMITED
IMPERIAL INNOVATIONS LIMITED
EMORY UNIVERSITY |
London, Greater London
London, Greater London
Atlanta |
GA |
GB
GB
US |
|
|
Family ID: |
50482596 |
Appl. No.: |
15/117982 |
Filed: |
February 20, 2015 |
PCT Filed: |
February 20, 2015 |
PCT NO: |
PCT/GB2015/050494 |
371 Date: |
August 10, 2016 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/519 20130101;
C07D 487/04 20130101; A61P 35/04 20180101 |
International
Class: |
C07D 487/04 20060101
C07D487/04 |
Foreign Application Data
Date |
Code |
Application Number |
Feb 21, 2014 |
GB |
1403093.6 |
Claims
1. A compound of the following formula: ##STR00206## or a
pharmaceutically acceptable salt or solvate thereof; wherein:
--R.sup.5X is -L.sup.5X-Q; -L.sup.5X- is independently a covalent
single bond or -L.sup.5XA-; -L.sup.5XA- is independently linear or
branched saturated C.sub.1-6alkylene, and is optionally substituted
with one or more groups selected from --OH and --OR.sup.L5X,
wherein each --R.sup.L5X is independently linear or branched
saturated C.sub.1-6alkyl or saturated C.sub.3-6cycloalkyl; -Q is a
non-aromatic heterocyclic ring having from 5 to 7 ring atoms,
including at least one nitrogen ring atom, and is substituted with
"n" groups -J, and is substituted with "m" groups --R.sup.Q; "n" is
1, 2, or 3; "m" is 0, 1, 2, or 3; each -J is independently --OH,
--OR.sup.J, -L.sup.J-OH, or -L.sup.J-OR.sup.J; each --R.sup.J is
independently linear or branched saturated C.sub.1-6alkyl or
saturated C.sub.3-6cycloalkyl; each -L.sup.J- is independently
linear or branched saturated C.sub.1-6alkylene; each --R.sup.Q is
independently independently --F, --Cl, --Br, --I, --R.sup.QA,
--CF.sub.3, --OCF.sub.3, --NH.sub.2, --NHR.sup.QA,
--NR.sup.QA.sub.2, pyrrolidino, piperidino, morpholino, piperazino,
N--(R.sup.QA)-piperazino, --SH, --SR.sup.QA, or --CN; each
--R.sup.QA is independently linear or branched saturated
C.sub.1-6alkyl or saturated C.sub.3-6cycloalkyl; --R.sup.5Y is
independently --H or --R.sup.5YA; --R.sup.5YA is
independentlylinear or branched saturated C.sub.1-6alkyl; --R.sup.7
is independently --R.sup.7X or --C(.dbd.O)R.sup.7X; each --R.sup.7X
is independently: --R.sup.7A, --R.sup.7B, --R.sup.7C, --R.sup.7D,
--R.sup.7E, -L.sup.7-R.sup.7B, -L.sup.7-R.sup.7C,
-L.sup.7-R.sup.7D, or -L.sup.7-R.sup.7E; each -L.sup.7- is
independently linear or branched saturated C.sub.1-6alkylene; each
--R.sup.7A is independently linear or branched saturated
C.sub.1-6alkyl, and is optionally substituted with one or more
substituents --W.sup.1; each --R.sup.7B is saturated
C.sub.3-6cycloalkyl, and is optionally substituted with one or more
substituents --W.sup.2; each --R.sup.7D is non-aromatic
C.sub.3-7heterocyclyl, and is optionally substituted with one or
more substituents --W.sup.2; each --R.sup.7D is phenyl or naphthyl,
and is optionally substituted with one or more substituents
--W.sup.3; each --R.sup.7E is C.sub.5-12heteroaryl, and is
optionally substituted with one or more substituents --W.sup.3;
each --W.sup.1 is independently: --F, --Cl, --Br, --I, --CF.sub.3,
--OH, --OR.sup.W1, --OCF.sub.3, --NH.sub.2, --NHR.sup.W1,
--NR.sup.W1.sub.2, pyrrolidino, piperidino, morpholino, piperazino,
N--(R.sup.W1)-piperazino, --C(.dbd.O)OH, --C(.dbd.O)OR.sup.W1,
--C(.dbd.O)NH.sub.2, --C(.dbd.O)NHR.sup.W1,
--C(.dbd.O)NR.sup.W1.sub.2, --C(.dbd.O)-pyrrolidino,
--C(.dbd.O)-piperidino, --C(.dbd.O)-morpholino,
--C(.dbd.O)-piperazino, --C(.dbd.O)--N--(R.sup.W1)-piperazino,
--S(.dbd.O)R.sup.W1, --S(.dbd.O).sub.2R.sup.W1,
--S(.dbd.O).sub.2NH.sub.2, --S(.dbd.O).sub.2NHR.sup.W1,
--S(.dbd.O).sub.2NR.sup.W1.sub.2, --S(.dbd.O).sub.2pyrrolidino,
--S(.dbd.O).sub.2-piperidino, --S(.dbd.O).sub.2-morpholino,
--S(.dbd.O).sub.2-piperazino,
--S(.dbd.O).sub.2--N--(R.sup.W1)-piperazino, --CN, or --NO.sub.2;
wherein each --R.sup.W1 is independently linear or branched
saturated C.sub.1-6alkyl, phenyl, or --CH.sub.2-phenyl, wherein
each phenyl is optionally substituted with one or more groups
selected from --F, --Cl, --Br, --I, --R.sup.W11, --CF.sub.3, --OH,
--OR.sup.W11, and --OCF.sub.3, wherein each --R.sup.W11 is
independently linear or branched saturated C.sub.1-6alkyl; each
--W.sup.2 is independently: --F, --Cl, --Br, --I, --R.sup.W2,
--CF.sub.3, --OH, --OR.sup.W2, --OCF.sub.3, --NH.sub.2,
--NHR.sup.W2, --NR.sup.W2.sub.2, pyrrolidino, piperidino,
morpholino, piperazino, N--(R.sup.W2)-piperazino, --C(.dbd.O)OH,
--C(.dbd.O)OR.sup.W2, --C(.dbd.O)NH.sub.2, --C(.dbd.O)NHR.sup.W2,
--C(.dbd.O)NR.sup.W2.sub.2, --C(.dbd.O)-pyrrolidino,
--C(.dbd.O)-piperidino, --C(.dbd.O)-morpholino,
--C(.dbd.O)-piperazino, --C(.dbd.O)--N--(R.sup.W2)-piperazino,
--S(.dbd.O)R.sup.W2, --S(.dbd.O).sub.2R.sup.W2,
--S(.dbd.O).sub.2NH.sub.2, --S(.dbd.O).sub.2NHR.sup.W2,
--S(.dbd.O).sub.2NR.sup.W2.sub.2, --S(.dbd.O).sub.2pyrrolidino,
--S(.dbd.O).sub.2-piperidino, --S(.dbd.O).sub.2-morpholino,
--S(.dbd.O).sub.2-piperazino,
--S(.dbd.O).sub.2--N--(R.sup.W2)-piperazino, --CN, or --NO.sub.2;
wherein each --R.sup.W2 is independently linear or branched
saturated C.sub.1-6alkyl, phenyl, or --CH.sub.2-phenyl, wherein
each phenyl is optionally substituted with one or more groups
selected from --F, --Cl, --Br, --I, --R.sup.W22, --CF.sub.3, --OH,
--OR.sup.W22, and --OCF.sub.3, wherein each --R.sup.W22 is
independently linear or branched saturated C.sub.1-6alkyl; each
--W.sup.3 is independently: --F, --Cl, --Br, --I, --R.sup.W3,
--CF.sub.3, --OH, --OR.sup.W3, --OCF.sub.3, --NH.sub.2,
--NHR.sup.W3, --NR.sup.W3.sub.2, pyrrolidino, piperidino,
morpholino, piperazino, N--(R.sup.W3)-piperazino, --C(.dbd.O)OH,
--C(.dbd.O)OR.sup.W3, --C(.dbd.O)NH.sub.2, --C(.dbd.O)NHR.sup.W3,
--C(.dbd.O)NR.sup.W3.sub.2, --C(.dbd.O)-pyrrolidino,
--C(.dbd.O)-piperidino, --C(.dbd.O)-morpholino,
--C(.dbd.O)-piperazino, --C(.dbd.O)--N--(R.sup.W3)-piperazino,
--S(.dbd.O)R.sup.W3, --S(.dbd.O).sub.2R.sup.W3,
--S(.dbd.O).sub.2NH.sub.2, --S(.dbd.O).sub.2NHR.sup.W3,
--S(.dbd.O).sub.2NR.sup.W3.sub.2, --S(.dbd.O).sub.2pyrrolidino,
--S(.dbd.O).sub.2-piperidino, --S(.dbd.O).sub.2-morpholino,
--S(.dbd.O).sub.2-piperazino,
--S(.dbd.O).sub.2--N--(R.sup.W3)-piperazino, --CN, or --NO.sub.2;
wherein each --R.sup.W3 is independently linear or branched
saturated C.sub.1-6alkyl, phenyl, or --CH.sub.2-phenyl, wherein
each phenyl is optionally substituted with one or more groups
selected from --F, --Cl, --Br, --I, --R.sup.W33, --CF.sub.3, --OH,
--OR.sup.W33, and --OCF.sub.3, wherein each --R.sup.W33 is
independently linear or branched saturated C.sub.1-6alkyl;
--R.sup.3 is independently --R.sup.3A or --R.sup.3B; --R.sup.3A is
independently linear or branched saturated C.sub.1-6alkyl;
--R.sup.3B is independently saturated C.sub.3-7cycloalkyl;
--R.sup.2 is independently --H or --R.sup.2A; --R.sup.2A is
independently --F, --Cl, --Br, --I, --R.sup.2AA, --CF.sub.3, --OH,
--OR.sup.2AA, --OCF.sub.3, --NH.sub.2, --NHR.sup.2AA,
--NR.sup.2AA.sub.2, pyrrolidino, piperidino, morpholino,
piperazino, N--(R.sup.2AA)-piperazino, --SH, --SR.sup.2AA, or --CN;
each --R.sup.2AA is independently linear or branched saturated
C.sub.1-6alkyl; --R.sup.6 is independently --H or --R.sup.6A;
--R.sup.6A is independently --F, --Cl, --Br, --I, --R.sup.6AA,
--CF.sub.3, --OH, --OR.sup.6AA, --OCF.sub.3, --NH.sub.2,
--NHR.sup.6AA, --NR.sup.6AA.sub.2, pyrrolidino, piperidino,
morpholino, piperazino, N--(R.sup.6AA)-piperazino, --SH,
--SR.sup.6AA, or --CN; and each --R.sup.6AAis independently linear
or branched saturated C.sub.1-6alkyl.
2. A compound according to claim 1, wherein -L.sup.5X- is a
covalent single bond.
3. A compound according to claim 1, wherein -L.sup.5X- is
-L.sup.5XA-.
4. A compound according to any one of claims 1 to 3, wherein
-L.sup.5XA-, if present, is independently linear or branched
saturated C.sub.1-4alkylene.
5. A compound according to any one of claims 1 to 3, wherein
-L.sup.5XA-, if present, is independently --CH.sub.2--,
--CH(CH.sub.3)--, --C(CH.sub.3).sub.2--, or
--CH.sub.2CH.sub.2--.
6. A compound according to any one of claims 1 to 3, wherein
-L.sup.5XA-, if present, is --CH.sub.2--.
7. A compound according to any one of claims 1 to 6, wherein -Q is
pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, azepanyl, or
diazepanyl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
8. A compound according to any one of claims 1 to 6, wherein -Q is
pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, azepanyl, or
diazepanyl, wherein the point of attachment is via a ring carbon
atom, and is substituted with "n" groups -J, and is substituted
with "m" groups --R.sup.Q.
9. A compound according to any one of claims 1 to 6, wherein -Q is
pyrrolidinyl, piperidinyl, morpholinyl, or piperazinyl, and is
substituted with "n" groups -J, and is substituted with "m" groups
--R.sup.Q.
10. A compound according to any one of claims 1 to 6, wherein -Q is
pyrrolidinyl, piperidinyl, morpholinyl, or piperazinyl, wherein the
point of attachment is via a ring carbon atom, and is substituted
with "n" groups -J, and is substituted with "m" groups
--R.sup.Q.
11. A compound according to any one of claims 1 to 6, wherein -Q is
pyrrolidinyl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
12. A compound according to any one of claims 1 to 6, wherein -Q is
pyrrolidinyl, wherein the point of attachment is via a ring carbon
atom, and is substituted with "n" groups -J, and is substituted
with "m" groups --R.sup.Q.
13. A compound according to any one of claims 1 to 6, wherein -Q is
pyrrolidin-2-yl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
14. A compound according to any one of claims 1 to 6, wherein -Q is
pyrrolidin-3-yl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
15. A compound according to any one of claims 1 to 6, wherein -Q is
piperidinyl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
16. A compound according to any one of claims 1 to 6, wherein -Q is
piperidinyl, wherein the point of attachment is via a ring carbon
atom, and is substituted with "n" groups -J, and is substituted
with "m" groups --R.sup.Q.
17. A compound according to any one of claims 1 to 6, wherein -Q is
piperidin-4-yl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
18. A compound according to any one of claims 1 to 6, wherein -Q is
piperidin-3-yl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
19. A compound according to any one of claims 1 to 18, wherein "n"
is 1.
20. A compound according to any one of claims 1 to 18, wherein "n"
is 2.
21. A compound according to any one of claims 1 to 20, wherein "m"
is 0.
22. A compound according to any one of claims 1 to 6, wherein -Q is
selected from: ##STR00207##
23. A compound according to any one of claims 1 to 6, wherein -Q
is: ##STR00208##
24. A compound according to any one of claims 1 to 6, wherein -Q
is: ##STR00209##
25. A compound according to any one of claims 1 to 6, wherein -Q
is: ##STR00210##
26. A compound according to any one of claims 1 to 6, wherein -Q is
selected from: ##STR00211##
27. A compound according to any one of claims 1 to 6, wherein -Q
is: ##STR00212##
28. A compound according to any one of claims 1 to 6, wherein -Q
is: ##STR00213##
29. A compound according to any one of claims 1 to 6, wherein -Q is
selected from: ##STR00214##
30. A compound according to any one of claims 1 to 6, wherein -Q
is: ##STR00215##
31. A compound according to any one of claims 1 to 6, wherein -Q
is: ##STR00216##
32. A compound according to any one of claims 1 to 6, wherein -Q
is: ##STR00217##
33. A compound according to any one of claims 1 to 6, wherein -Q is
selected from: ##STR00218##
34. A compound according to any one of claims 1 to 6, wherein -Q
is: ##STR00219##
35. A compound according to any one of claims 1 to 6, wherein -Q
is: ##STR00220##
36. A compound according to any one of claims 1 to 6, wherein -Q
is: ##STR00221##
37. A compound according to any one of claims 1 to 36, wherein each
-J is independently --OH or -La-OH.
38. A compound according to any one of claims 1 to 36, wherein each
-J is --OH.
39. A compound according to any one of claims 1 to 36, wherein each
-J is --OR.sup.J.
40. A compound according to any one of claims 1 to 36, wherein each
-J is -L.sup.J-OH.
41. A compound according to any one of claims 1 to 40, wherein each
--R.sup.J, if present, is independently -Me, -Et, -nPr, -iPr, -nBu,
-iBu, -sBu, or -tBu.
42. A compound according to any one of claims 1 to 40, wherein each
--R.sup.J, if present, is -Me.
43. A compound according to any one of claims 1 to 42, wherein each
-L.sup.J-, if present, is independently --CH.sub.2--,
--CH(CH.sub.3)--, --C(CH.sub.3).sub.2--, --CH.sub.2CH.sub.2--,
--CH(CH.sub.3)CH.sub.2--, --CH.sub.2CH(CH.sub.3)-, or
--CH.sub.2CH.sub.2CH.sub.2--.
44. A compound according to any one of claims 1 to 42, wherein each
-L.sup.J-, if present, is independently --CH.sub.2--.
45. A compound according to any one of claims 1 to 44, wherein
--R.sup.5Y is --H.
46. A compound according to any one of claims 1 to 45, wherein
--R.sup.7 is --R.sup.7X.
47. A compound according to any one of claims 1 to 45, wherein
--R.sup.7 is --C(.dbd.O)R.sup.7X.
48. A compound according to any one of claims 1 to 47, wherein each
--R.sup.7X is independently: --R.sup.7C, --R.sup.7D, --R.sup.7E,
-L.sup.7-R.sup.7B, -L.sup.7-R.sup.7D, or -L.sup.7-R.sup.7E.
49. A compound according to any one of claims 1 to 47, wherein each
--R.sup.7X is independently: -L.sup.7R.sup.7B, -L.sup.7-R.sup.7D,
or -L.sup.7-R.sup.7E.
50. A compound according to any one of claims 1 to 47, wherein each
--R.sup.7X is --R.sup.7D.
51. A compound according to any one of claims 1 to 47, wherein each
--R.sup.7X is --R.sup.7D.
52. A compound according to any one of claims 1 to 47, wherein each
--R.sup.7X is --R.sup.7E.
53. A compound according to any one of claims 1 to 47, wherein each
--R.sup.7X is -L.sup.7-R.sup.7B.
54. A compound according to any one of claims 1 to 47, wherein each
--R.sup.7X is -L.sup.7-R.sup.7D.
55. A compound according to any one of claims 1 to 47, wherein each
--R.sup.7X is -L.sup.7-R.sup.7E.
56. A compound according to any one of claims 1 to 55, wherein each
-L.sup.7-, if present, is independently --CH.sub.2--,
--CH(CH.sub.3)--, --C(CH.sub.3).sub.2--, --CH.sub.2CH.sub.2--,
--CH(CH.sub.3)CH.sub.2--, --CH.sub.2CH(CH.sub.3)--, or
--CH.sub.2CH.sub.2CH.sub.2--.
57. A compound according to any one of claims 1 to 55, wherein each
-L.sup.7-, if present, is --CH.sub.2--.
58. A compound according to any one of claims 1 to 57, wherein each
--R.sup.7E, if present, is independently cyclopropyl, cyclobutyl,
cyclopentyl, or cyclohexyl, and is optionally substituted with one
or more substituents --W.sup.2.
59. A compound according to any one of claims 1 to 57, wherein each
--R.sup.7E, if present, is cyclohexyl, and is optionally
substituted with one or more substituents --W.sup.2.
60. A compound according to any one of claims 1 to 57, wherein each
--R.sup.7E, if present, is cyclohexyl.
61. A compound according to any one of claims 1 to 60, wherein each
--R.sup.7D, if present, is independently pyrrolidinyl, piperidinyl,
morpholinyl, piperazinyl, tetrahydrofuranyl, tetrahydropyranyl,
dixoanyl, azepanyl, or diazepanyl, and is optionally substituted
with one or more substituents --W.sup.2.
62. A compound according to any one of claims 1 to 60, wherein each
--R.sup.7D, if present, is independently pyrrolidinyl, piperidinyl,
morpholinyl, or piperazinyl, and is optionally substituted with one
or more substituents --W.sup.2.
63. A compound according to any one of claims 1 to 60, wherein each
--R.sup.7D, if present, is independently piperidinyl, and is
optionally substituted with one or more substituents --W.sup.2.
64. A compound according to any one of claims 1 to 60, wherein each
--R.sup.7D, if present, is independently piperidinyl.
65. A compound according to any one of claims 1 to 64, wherein each
--R.sup.7D, if present, is phenyl, and is optionally substituted
with one or more substituents --W.sup.3.
66. A compound according to any one of claims 1 to 64, wherein each
--R.sup.7D, if present, is phenyl.
67. A compound according to any one of claims 1 to 66, wherein each
--R.sup.7E, if present, is C.sub.5-6heteroaryl, and is optionally
substituted with one or more substituents --W.sup.3.
68. A compound according to any one of claims 1 to 66, wherein each
--R.sup.7E, if present, is C.sub.9-10heteroaryl, and is optionally
substituted with one or more substituents --W.sup.3.
69. A compound according to any one of claims 1 to 66, wherein each
--R.sup.7E, if present, is independently furanyl, thienyl,
pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl,
isothiazolyl, triazolyl (e.g., 1H-[1,2,3]triazolyl,
2H-[1,2,3]triazolyl, 4H-[1,2,4]triazolyl, 1H-[1,2,4]triazolyl),
oxadiazolyl (e.g., [1,2,3]oxadiazolyl, furazanyl,
[1,3,4]oxadiazolyl, [1,2,4]oxadiazolyl), thiadiazolyl (e.g.,
[1,2,3]thiadiazolyl, [1,2,5]thiadiazolyl, [1,3,4]thiadiazolyl,
[1,2,4]thiadiazolyl), tetrazolyl, pyridyl, pyrimidinyl, pyrazinyl,
pyridazinyl, or triazinyl (e.g., [1,3,5]-triazinyl), and is
optionally substituted with one or more substituents --W.sup.3.
70. A compound according to any one of claims 1 to 66, wherein each
--R.sup.7E, if present, is independently pyridyl, pyrimidinyl,
pyrazinyl, or pyridazinyl, and is optionally substituted with one
or more substituents --W.sup.3.
71. A compound according to any one of claims 1 to 66, wherein each
--R.sup.7E, if present, is pyridyl, and is optionally substituted
with one or more substituents --W.sup.3.
72. A compound according to any one of claims 1 to 66, wherein each
--R.sup.7E, if present, is pyridyl.
73. A compound according to any one of claims 1 to 66, wherein each
--R.sup.7E, if present, is independently indolyl, indazolyl,
benzofuranyl, benzothienyl, benzimidazolyl, benzothiazolyl,
benzoxazolyl, benzoisoxazolyl, quinolinyl, isoquinolinyl,
cinnolinyl, quinazolinyl, phthalazinyl, or quinoxalinyl, and is
optionally substituted with one or more substituents --W.sup.3.
74. A compound according to any one of claims 1 to 66, wherein each
--R.sup.7E, if present, is benzothiazolyl, and is optionally
substituted with one or more substituents --W.sup.3.
75. A compound according to any one of claims 1 to 66, wherein each
--R.sup.7E, if present, is benzothiazolyl.
76. A compound according to any one of claims 1 to 75, wherein each
--W.sup.2, if present, is independently: --F, --Cl, --Br, --I,
--CF.sub.3, --OH, --R.sup.W2, --OCF.sub.3, --NH.sub.2,
--NHR.sup.W2, --NR.sup.W2.sub.2, pyrrolidino, piperidino,
morpholino, piperazino, or N--(R.sup.W2)-piperazino.
77. A compound according to any one of claims 1 to 75, wherein each
--W.sup.2, if present, is independently: --F, --Cl, --Br, --I,
--CF.sub.3, --OH, --OR.sup.W2, or --OCF.sub.3.
78. A compound according to any one of claims 1 to 77, wherein each
--R.sup.W2, if present, is independently linear or branched
saturated C.sub.1-4alkyl, phenyl, or --CH.sub.2-phenyl.
79. A compound according to any one of claims 1 to 77, wherein each
--R.sup.W2, if present, is independently linear or branched
saturated C.sub.1-4alkyl.
80. A compound according to any one of claims 1 to 77, wherein each
--R.sup.W2, if present, is -Me.
81. A compound according to any one of claims 1 to 80, wherein each
--W.sup.3, if present, is independently: --F, --Cl, --Br, --I,
--CF.sub.3, --OH, --R.sup.W3, --OCF.sub.3, --NH.sub.2,
--NHR.sup.W3, --NR.sup.W3.sub.2, pyrrolidino, piperidino,
morpholino, piperazino, or N--(R.sup.W3)-piperazino.
82. A compound according to any one of claims 1 to 80, wherein each
--W.sup.3, if present, is independently: --F, --Cl, --Br, --I,
--CF.sub.3, --OH, --OR.sup.W3, or --OCF.sub.3.
83. A compound according to any one of claims 1 to 82, wherein each
--R.sup.W3, if present, is independently linear or branched
saturated C.sub.1-4alkyl, phenyl, or --CH.sub.2-phenyl.
84. A compound according to any one of claims 1 to 82, wherein each
--R.sup.W3, if present, is independently linear or branched
saturated C.sub.1-4alkyl.
85. A compound according to any one of claims 1 to 82, wherein each
--R.sup.W3, if present, is -Me.
86. A compound according to any one of claims 1 to 85, wherein
--R.sup.3 is --R.sup.3A.
87. A compound according to any one of claims 1 to 85, wherein
--R.sup.3 is --R.sup.3B.
88. A compound according to any one of claims 1 to 87, wherein
--R.sup.3A, if present, is independently -Me, -Et, -nPr, -iPr,
-nBu, -iBu, -sBu, or -tBu, n-pentyl, t-pentyl, neo-pentyl,
iso-pentyl, sec-pentyl, 3-pentyl, 1-hexyl, 2-hexyl, 3-hexyl,
3-methyl-1-pentyl, 4-methyl-1-pentyl, 4-methyl-2-pentyl,
4-methyl-3-pentyl, 2-methyl-2-pentyl, 2-methyl-1-pentyl,
2-methyl-2-pentyl, 3,3-dimethyl-1-butyl, 3,3-dimethyl-2-butyl,
3-methyl-1-pentyl, 3-methyl-2-pentyl, 3-methyl-3-pentyl,
2,2-dimethyl-1-butyl, 2,3-dimethyl-1-butyl, or
2,3-dimethyl-2-butyl.
89. A compound according to any one of claims 1 to 87, wherein
--R.sup.3A, if present, is independently -Me, -Et, -nPr, -iPr,
-nBu, -iBu, -sBu, or -tBu.
90. A compound according to any one of claims 1 to 87, wherein
--R.sup.3A, if present, is -iPr.
91. A compound according to any one of claims 1 to 90, wherein
--R.sup.3B, if present, is independently cyclopropyl, cyclobutyl,
cyclopentyl, or cyclohexyl.
92. A compound according to any one of claims 1 to 91, wherein
--R.sup.2 is --H.
93. A compound according to any one of claims 1 to 92, wherein
--R.sup.6 is --H.
94. A compound according to claim 1, selected from compounds of the
following formulae and pharmaceutically acceptable salts, hydrates,
and solvates thereof: PPDA-001 through PPDA-032.
95. A composition comprising a compound according to any one of
claims 1 to 94, and a pharmaceutically acceptable carrier or
diluent.
96. A method of preparing a composition comprising the step of
mixing a compound according to any one of claims 1 to 94, and a
pharmaceutically acceptable carrier or diluent.
97. A method of inhibiting CDK function in a cell, in vitro or in
vivo, comprising contacting the cell with an effective amount of a
compound according to any one of claims 1 to 94.
98. A method of regulating (e.g., inhibiting) cell proliferation
(e.g., proliferation of a cell), inhibiting cell cycle progression,
promoting apoptosis, or a combination of one or more these, in
vitro or in vivo, comprising contacting a cell with an effective
amount of a compound according to any one of claims 1 to 94.
99. A compound according to any one of claims 1 to 94, for use in a
method of treatment of the human or animal body by therapy.
100. A compound according to any one of claims 1 to 94, for use in
a method of treatment of a disorder.
101. Use of a compound according to any one of claims 1 to 94, in
the manufacture of a medicament for use in a method of treatment of
a disorder.
102. A method of treatment of a disorder, comprising administering
to a subject in need of treatment a therapeutically-effective
amount of a compound according to any one of claims 1 to 94.
103. A compound for use according to claim 100, use according to
claim 101, or a method according to claim 102, wherein the disorder
is: a disorder that is associated with CDK; a disorder resulting
from an inappropriate activity of a cyclin-dependent kinase (CDK);
a disorder that is associated with CDK mutation; a disorder that is
associated with CDK overexpression; a disorder that is associated
with upstream pathway activation of CDK; or a disorder that is
ameliorated by the inhibition of CDK.
104. A compound for use according to claim 100, use according to
claim 101, or a method according to claim 102, wherein the disorder
is: a proliferative disorder; cancer; a viral infection (e.g.,
HIV); a neurodegenerative disorder (e.g., Alzheimer's disease,
Parkinson's disease); ischaemia; a renal disease; or a
cardiovascular disorder (e.g., atherosclerosis).
105. A compound for use according to claim 100, use according to
claim 101, or a method according to claim 102, wherein the disorder
is: a proliferative disorder.
106. A compound for use according to claim 100, use according to
claim 101, or a method according to claim 102, wherein the disorder
is: cancer.
107. A compound for use according to any one of claims 100 and 103
to 106, use according to any one of claims 101 and 103 to 106, or a
method according to any one of claims 102 to 106, wherein the
treatment further comprises treatment (e.g., simultaneous or
sequential treatment) with a further active agent which is an
aromatase inhibitor.
108. A compound for use according to claim 107, use according to
claim 107, or a method according to claim 107, wherein the
aromatase inhibitor is exemestane (also known as Aromasin),
letrozole (also known as Femara), or anastrozole (also known as
Arimidex).
109. A compound for use according to claim 107 or 108, use
according to 107 or 108, or a method according to claim 107 or 108,
wherein the disorder is breast cancer (e.g., breast cancer which is
resistant to said aromatase inhibitor).
110. A compound for use according to any one of claims 100 and 103
to 106, use according to any one of claims 101 and 103 to 106, or a
method according to any one of claims 102 to 106, wherein the
treatment further comprises treatment (e.g., simultaneous or
sequential treatment) with a further active agent which is an
anti-estrogen.
111. A compound for use according to claim 110, use according to
claim 110, or a method according to claim 110, wherein the
anti-estrogen is faslodex (also known as Fulvestrant and
ICI182780), tamoxifen (also known as Nolvadex), or
hydroxytamoxifen.
112. A compound for use according to claim 110 or 111, use
according to 110 or 111, or a method according to 110 or 111,
wherein the disorder is breast cancer (e.g., breast cancer which is
resistant to said anti-estrogen).
113. A compound for use according to any one of claims 100 and 103
to 106, use according to any one of claims 101 and 103 to 106, or a
method according to any one of claims 102 to 106, wherein the
treatment further comprises treatment (e.g., simultaneous or
sequential treatment) with a further active agent which is a Her2
blocker.
114. A compound for use according to claim 113, use according to
claim 113, or a method according to claim 113, wherein the Her2
blocker is herceptin, pertuzumab, or lapatinib.
115. A compound for use according to any one of claims 100 and 103
to 106, use according to any one of claims 101 and 103 to 106, or a
method according to any one of claims 102 to 106, wherein the
treatment further comprises treatment (e.g., simultaneous or
sequential treatment) with a further cytotoxic chemotherapeutic
agent.
116. A compound for use according to claim 115, use according to
claim 115, or a method according to claim 115, wherein the
cytotoxic chemotherapeutic agent is a taxane (e.g., paclitaxel also
known as Taxol; docetaxel also known as Taxotere),
cyclophosphamide, or an antimetabolite (e.g., carboplatin,
capecitabine, gemcitabine, doxorubicin, epirubicin, 5-fluorouracil,
etc.).
Description
RELATED APPLICATION
[0001] This application is related to United Kingdom patent
application number 1403093.6 filed 21 Feb. 2014, the contents of
which are incorporated herein by reference in their entirety.
TECHNICAL FIELD
[0002] The present invention pertains generally to the field of
therapeutic compounds. More specifically the present invention
pertains to certain pyrazolo[1,5-a]pyrimidine-5,7-diamine compounds
(referred to herein as "PPDA compounds") that, inter alia, inhibit
(e.g., selectively inhibit) CDK (e.g., CDK1, CDK2, CDK4, CDK5,
CDK6, CDK7, CDK8, CDK9, CDK10, CDK11, CDK12, CDK13, etc.). The
present invention also pertains to pharmaceutical compositions
comprising such compounds, and the use of such compounds and
compositions, both in vitro and in vivo, to inhibit CDK; and to
treat disorders including: disorders that are associated with CDK;
disorders that result from an inappropriate activity of a
cyclin-dependent kinase (CDK); disorders that are associated with
CDK mutation; disorders that are associated with CDK
overexpression; disorders that are associated with upstream pathway
activation of CDK; disorders that are ameliorated by the inhibition
of CDK; proliferative disorders; cancer; viral infections
(including HIV); neurodegenerative disorders (including Alzheimer's
disease and Parkinson's disease); ischaemia; renal diseases; and
cardiovascular disorders (including atherosclerosis). Optionally,
the treatment further comprises treatment (e.g., simultaneous or
sequential treatment) with a further active agent which is, e.g.,
an aromatase inhibitor, an anti-estrogen, a Her2 blocker, a
cytotoxic chemotherapeutic agent, etc.
BACKGROUND
[0003] A number of publications are cited herein in order to more
fully describe and disclose the invention and the state of the art
to which the invention pertains. Each of these references is
incorporated herein by reference in its entirety into the present
disclosure, to the same extent as if each individual reference was
specifically and individually indicated to be incorporated by
reference.
[0004] Throughout this specification, including the claims which
follow, unless the context requires otherwise, the word "comprise,"
and variations such as "comprises" and "comprising," will be
understood to imply the inclusion of a stated integer or step or
group of integers or steps but not the exclusion of any other
integer or step or group of integers or steps.
[0005] It must be noted that, as used in the specification and the
appended claims, the singular forms "a," "an," and "the" include
plural referents unless the context clearly dictates otherwise.
Thus, for example, reference to "a pharmaceutical carrier" includes
mixtures of two or more such carriers, and the like.
[0006] Ranges are often expressed herein as from "about" one
particular value, and/or to "about" another particular value. When
such a range is expressed, another embodiment includes from the one
particular value and/or to the other particular value. Similarly,
when values are expressed as approximations, by the use of the
antecedent "about," it will be understood that the particular value
forms another embodiment.
[0007] This disclosure includes information that may be useful in
understanding the present invention. It is not an admission that
any of the information provided herein is prior art or relevant to
the presently claimed invention, or that any publication
specifically or implicitly referenced is prior art.
[0008] Cyclin-Dependent Protein Kinase (CDK)
[0009] Cyclin-dependent protein kinases (CDK) are the catalytic
subunits of a family of 21 serine/threonine protein kinases (see,
e.g., Malumbres et al., 2009), some of which control progression of
the cell through the stages of growth, DNA replication and mitosis
(see, e.g., Pines, 1995; Morgan, 1995). Activation of specific CDKs
is required for appropriate progression through the different
stages of the cell cycle and entry into the next stage of the cell
cycle. CDK4 and CDK6 are required for progression through the
growth (G1) phase, CDK2 in the DNA synthesis (S phase) and CDK1 for
mitosis and cell division (M phase). Regulation of the activity of
the cell cycle CDKs is pivotal for correct timing of progression of
the cell through the stages of the cell cycle and their activities
are regulated at many levels, including complex formation with
specific cyclins (A, B, D and E class cyclins; these cyclins are
synthesized and degraded through the stages of the cell cycle), CDK
inhibitors (CDKI), in particular CIP/KIP and INK-type CDKIs (see,
e.g., Sherr et al., 1995), as well as phosphorylation and
dephosphorylation at specific residues. The phosphorylation status
of a specific threonine residue in the activation loop, the
so-called T-loop, is a key modification for the activity of cell
cycle CDKs (see, e.g., Fisher et al., 1994).
[0010] De-regulation of CDK activity is an important component of
many disease states, generally through elevated and/or
inappropriate activation, as CDKs themselves are infrequently
mutated. Rare examples of mutations in cell cycle CDKs include CDK4
families with hereditary melanoma that result in insensitivity to
the INK4 CDKIs (see, e.g., Zuo et al, 1996). Inactivating mutations
in the CDKN2A gene, which encodes for p16INK4 and p14ARF CDKIs, are
more common in hereditary melanoma (see, e.g., Hansson, 2010),
these mutations also being associated with greater incidence of
breast and pancreatic cancer in affected families (see, e.g., Borg
et al., 2000). CDK4 and CDK6 can be amplified and/or overexpressed
in cancer, their cyclin effectors, D-type cyclins, are also often
amplified and/or over-expressed, whilst the CDK4/CDK6 inhibitors
(INK4 genes) are frequently deleted in many cancer types and/or
undergo epigenetic silencing (see, e.g., Ortega et al., 2002).
E-type cyclins interact with CDK2 for its activity and are
frequently over-expressed in cancer, whilst the p21 and p27
inhibitory proteins that act on CDK2, as well as CDK1, are
epigenetically silenced in cancer (see, e.g., Malumbres et al.,
2001; Jones et al., 2007). Up-regulation of the activities of cell
cycle CDKs is therefore integral to cancer development and
progression.
[0011] CDK7, another member of the CDK family, which complexes with
cyclin H and MAT1, phosphorylates the cell cycle CDKs in the
activation of T-loop, to promote their activities (see, e.g.,
Fisher et al., 1994). As such, it has been proposed that inhibiting
CDK7 would provide a potent means of inhibiting cell cycle
progression, which may be especially relevant given that there is
compelling evidence from gene knockout studies in mice for lack of
an absolute requirement for CDK2, CDK4 and CDK6 for the cell cycle,
at least in most cell types (see, e.g., Malumbres et al., 2009),
whilst different tumors appear to require some, but be independent
of other interphase CDKs (CDK2, CDK4, CDK6). Recent genetic and
biochemical studies have confirmed the importance of CDK7 for cell
cycle progression (see, e.g., Larochelle et al., 2007; Ganuza et
al., 2012).
[0012] In addition to its role as the CDK Activating Kinase (CAK),
CDK7/cyclin H/MAT1, in complex with the basal transcription factor
TFIIH, phosphorylates RNA polymerase II (PoIII) in its C-terminal
domain (CTD) (see, e.g., Lu et al., 1995; Serizawa et al., 1995).
CDK9, another member of the family, is also required for PoIII CTD
phosphorylation. The PoIII CTD is comprised of a seven amino acid
repeat having the sequence
Tyrosine-Serine-Proline-Threonine-Serine-Proline-Serine (YSPTSPS),
52 YSPTSPS heptad repeats being present in the mammalian PoIII CTD.
Phosphorylation of serine-2 (S2) and serine-5 (S5) by CDK7 and CDK9
is required for release of PoIII from the gene promoter at
initiation of transcription. CDK7 appears to act upstream of CDK9,
phosphorylation of S5 phosphorylation by CDK7 preceding S2
phosphorylation by CDK9 (see, e.g., Larochelle et al., 2012).
Transcriptional inhibitors such as flavopiridol, as well as CDK
inhibitors that inhibit CDK7 and CDK9 demonstrate the potential
utility of CDK7 and CDK9 inhibition in cancer (see, e.g., Wang et
al., 2008). In addition to their action in phosphorylating the
PoIII CTD, CDK7 and CDK9 have been implicated in regulating the
activities of a number of transcription factors, including the
breast cancer associated estrogen receptor (ER) (see, e.g., Chen et
al., 2000), retinoid receptors (see, e.g., Rochette-Egly et al.,
1997; Bastien et al., 2000), the androgen receptor (see, e.g.,
Chymkowitch et al., 2011; Gordon et al., 2010), as well as the
tumor suppressor p53 (Lu et al., 1997; Ko et al., 1997;
Radhakrishnan et al., 2006; Claudio et al., 2006). CDK8, a
component of the mediator complex that regulates gene
transcription, through a mechanism involving interaction between
transcription factors and the PoIII basal transcription machinery,
also phosphorylates transcription factors to regulate their
activities (see, e.g., Alarcon et al., 2009). CDK8 also appears to
be important for regulating transcription reinitiation. The
importance of CDK8 in cancer is highlighted by the finding that the
CDK8 gene is amplified in 40-60% of colorectal cancers, whilst its
cyclin partner, cyclin c, is upregulated in many cancer types,
whilst functional studies are supportive of an oncogenic role for
CDK8 in cancer (see, e.g., Xu et al., 2011). A potential role for
CDK11 in regulating mediator activity has been described,
indicating a role for CDK11 in transcription regulation (see, e.g.,
Drogat et al., 2012), whilst their ability to phosphorylate S2 the
PoIII CTD also implicates CDK12 and CDK13 in transcription; CDK12
is also implicated in maintenance of genome stability (see, e.g.,
Bartkowiak et al., 2010; Blazek et al., 2011; Cheng et al.,
2012).
[0013] In addition to the great deal of evidence implicating the
above and other CDKs (e.g., CDK10; see, e.g., Lorns et al., 2008;
Yu et al., 2012) in cancer, CDKs are also important in viral
infections including HIV (see, e.g., Knockeart et al., 2002),
neurodegenerative disorders including Alzheimer's and Parkinson's
disease (of particular note here is CDK5, see, e.g., Monaco et al.,
2005; Faterna et al., 2008), ischaemia, and proliferative
disorders, including renal diseases (see, e.g., Marshall et al.,
2006) and cardiovascular disorders including atherosclerosis.
[0014] The development of small molecule CDK inhibitors provides a
potentially powerful approach in the treatment of many human
diseases, in particular cancer. Thus inhibition of cell cycle
progression may be achieved through the development of selective
CDK1 inhibitors (as CDK1 appears to be indispensible for the cell
cycle) or selective CDK7 inhibitors (as CDK7 regulates the cell
cycle CDKs) or with inhibitors with activity against all of the
cell cycle CDKs. Some evidence indicates that selective CDK4/CDK6
or CDK2 inhibitors may have utility for specific conditions (e.g.,
CDK4/CDK6 in haematological malignancies and CDK2 in glioblastomas
or osteosarcomas), and so development of selective inhibitors for
these CDKs may be of utility, the selectivity perhaps aiding
toxicity issues.
[0015] Known Compounds
[0016] It appears that the following compounds are known.
TABLE-US-00001 CAS Registry No. Structure 771502-87-5 ##STR00001##
771501-59-8 ##STR00002## 771509-61-6 ##STR00003## 771502-45-5
##STR00004## 771508-20-4 ##STR00005## 1092443-65-6 ##STR00006##
1092443-63-4 ##STR00007## 1092444-59-1 ##STR00008## 1092444-58-0
##STR00009## 1092444-23-9 ##STR00010## 1092444-03-5 ##STR00011##
1256288-39-7 ##STR00012##
BRIEF DESCRIPTION OF THE DRAWINGS
[0017] FIG. 1 is a graph of percentage growth inhibition as a
function of the base-10 logarithm of the molar concentration of the
test compound, PPDA-001, as determined by the NCI60 cancer cell
line screen. Each line represents one cell line.
[0018] FIG. 2 is a graph of relative tumour volume as a function of
time in the HCT116 tumour xenograft study, for vehicle control
(squares), 50 mg/kg/bi-daily (triangles), and 100 mg/kg once daily
(crosses). Error bars represent standard errors of the mean
(SEM).
[0019] FIG. 3 is a graph of percent body weight as a function of
time in the HCT116 tumour xenograft study, for vehicle control
(squares), 50 mg/kg/bi-daily (triangles), and 100 mg/kg once daily
(crosses).
[0020] FIG. 4, which is a graph of relative growth (mean
growth.+-.standard errors of the mean) as function of time for
treatment with: (a) vehicle; (b) PPDA-001/ICEC0942; (c)
4-hydroxytamoxifen; and (d) PPDA-001/ICEC0942 with
4-hydroxytamoxifen. As shown, co-treatment greatly enhances the
growth inhibition observed for each drug alone.
[0021] FIG. 5, which is a graph of relative growth (mean
growth.+-.standard errors of the mean) as function of time for
treatment with: (a) vehicle; (b) PPDA-001/ ICEC0942; (c) Faslodex;
and (d) PPDA-001/ICEC0942 with Faslodex. As shown, co-treatment
greatly enhances the growth inhibition observed for each drug
alone.
[0022] FIG. 6 shows immunoblot gels for the bands for
phospho-ER.sup.S118, ER, and .beta.-actin, as a function of time,
for cell lysates from MCF-7 cells treated with 1 .mu.mol/L
PPDA-001/ICEC0942. As shown in the figure, PPDA-001/ICEC0942
inhibits phosphorylation of ER at serine-118
(phospho-ER.sup.S118).
[0023] FIG. 7 shows immunoblot gels for cell lysates from MCF-7
cells treated for 24 hours with 10 .mu.mol/L PPDA-001/ICEC0942.
SUMMARY OF THE INVENTION
[0024] One aspect of the invention pertains to certain certain
pyrazolo[1,5-a]pyrimidine-5,7-diamine compounds (referred to herein
as "PPDA compounds"), as described herein.
[0025] Another aspect of the invention pertains to a composition
(e.g., a pharmaceutical composition) comprising a PPDA compound, as
described herein, and a pharmaceutically acceptable carrier or
diluent.
[0026] Another aspect of the invention pertains to a method of
preparing a composition (e.g., a pharmaceutical composition)
comprising the step of mixing a PPDA compound, as described herein,
and a pharmaceutically acceptable carrier or diluent.
[0027] Another aspect of the present invention pertains to a method
of inhibiting CDK (e.g., CDK1, CDK2, CDK4, CDK5, CDK6, CDK7, CDK8,
CDK9, CDK10, CDK11, CDK12, CDK13, etc.) function (e.g., in a cell),
in vitro or in vivo, comprising contacting the cell with an
effective amount of a PPDA compound, as described herein.
[0028] Another aspect of the present invention pertains to a method
of regulating (e.g., inhibiting) cell proliferation (e.g.,
proliferation of a cell), inhibiting cell cycle progression,
promoting apoptosis, or a combination of one or more these, in
vitro or in vivo, comprising contacting a cell with an effective
amount of a PPDA compound, as described herein.
[0029] Another aspect of the present invention pertains to a PPDA
compound as described herein for use in a method of treatment of
the human or animal body by therapy, for example, for use a method
of treatment of a disorder (e.g., a disease) as described
herein.
[0030] Another aspect of the present invention pertains to use of a
PPDA compound, as described herein, in the manufacture of a
medicament, for example, for use in a method of treatment, for
example, for use a method of treatment of a disorder (e.g., a
disease) as described herein.
[0031] Another aspect of the present invention pertains to a method
of treatment, for example, a method of treatment of a disorder
(e.g., a disease) as described herein, comprising administering to
a subject in need of treatment a therapeutically-effective amount
of a PPDA compound, as described herein, preferably in the form of
a pharmaceutical composition.
[0032] In one embodiment, the treatment further comprises treatment
(e.g., simultaneous or sequential treatment) with a further active
agent which is, e.g., an aromatase inhibitor, an anti-estrogen, a
Her2 blocker, a cytotoxic chemotherapeutic agent, etc., as
described herein.
[0033] Another aspect of the present invention pertains to a kit
comprising (a) a PPDA compound, as described herein, preferably
provided as a pharmaceutical composition and in a suitable
container and/or with suitable packaging; and (b) instructions for
use, for example, written instructions on how to administer the
compound.
[0034] Another aspect of the present invention pertains to a PPDA
compound obtainable by a method of synthesis as described herein,
or a method comprising a method of synthesis as described
herein.
[0035] Another aspect of the present invention pertains to a PPDA
compound obtained by a method of synthesis as described herein, or
a method comprising a method of synthesis as described herein.
[0036] Another aspect of the present invention pertains to novel
intermediates, as described herein, which are suitable for use in
the methods of synthesis described herein.
[0037] Another aspect of the present invention pertains to the use
of such novel intermediates, as described herein, in the methods of
synthesis described herein.
[0038] As will be appreciated by one of skill in the art, features
and preferred embodiments of one aspect of the invention will also
pertain to other aspects of the invention.
DETAILED DESCRIPTION OF THE INVENTION
[0039] Compounds
[0040] One aspect of the present invention relates to certain
compounds which are related to
pyrazolo[1,5-a]pyrimidine-5,7-diamine:
##STR00013##
[0041] All of the compounds of the present invention have:
[0042] (a) a substituted amino group at the 5-position (denoted
herein as --NR.sup.5XR.sup.6Y);
[0043] (b) a substituted amino group at the 7-position (denoted
herein as --NHR.sup.7); and
[0044] (c) an alkyl or cycloalkyl group at the 3-position (denoted
herein as --R.sup.3).
[0045] More specifically, the group --R.sup.5X is, or contains, a
non-aromatic heterocyclic ring having from 5 to 7 ring atoms,
including at least one nitrogen ring atom (denoted herein as -Q),
and substituted with at least one "oxy" substituent (denoted herein
as -J).
[0046] Thus, one aspect of the present invention is a compound of
the following formula, or a pharmaceutically acceptable salt,
hydrate, or solvate thereof, wherein --R.sup.2, --R.sup.3,
--R.sup.5X, --R.sup.5Y, --R.sup.6, and --R.sup.7 are as defined
herein (for convenience, collectively referred to herein as
"pyrazolo[1,5-a]pyrimidine-5,7-diamine compounds" and "PPDA
compounds"):
##STR00014##
[0047] Some embodiments of the invention include the following:
[0048] (1) A compound of the following formula:
##STR00015##
[0049] or a pharmaceutically acceptable salt or solvate
thereof;
[0050] wherein: [0051] --R.sup.5X is -L.sup.5X-Q; [0052] -L.sup.5X-
is independently a covalent single bond or -L.sup.5XA-; [0053]
-L.sup.5XA- is independently linear or branched saturated
C.sub.1-6alkylene, and is optionally substituted with one or more
groups selected from --OH and --OR.sup.L5X, wherein each
--R.sup.L5X is independently linear or branched saturated
C.sub.1-6alkyl or saturated C.sub.3-6cycloalkyl; [0054] -Q is a
non-aromatic heterocyclic ring having from 5 to 7 ring atoms,
including at least one nitrogen ring atom, and is substituted with
"n" groups -J, and is substituted with "m" groups --R.sup.Q; [0055]
"n" is 1, 2, or 3; [0056] "m" is 0, 1, 2, or 3; [0057] each -J is
independently --OH, --OR.sup.J, -L.sup.J-OH, or -L.sup.J-OR.sup.J;
[0058] each --R.sup.J is independently linear or branched saturated
C.sub.1-6alkyl or saturated C.sub.3-6cycloalkyl; [0059] each
-L.sup.J- is independently linear or branched saturated
C.sub.1-6alkylene; [0060] each --R.sup.Q is independently --F,
--Cl, --Br, --I, --R.sup.QA, --CF.sub.3, --OCF.sub.3, --NH.sub.2,
--NHR.sup.QA, --NR.sup.QA.sub.2, pyrrolidino, piperidino,
morpholino, piperazino, N-(R.sup.QA)-piperazino, --SH, --SR.sup.QA,
or --CN; [0061] each --R.sup.QA is independently linear or branched
saturated C.sub.1-6alkyl or saturated C.sub.3-6cycloalkyl; [0062]
--R.sup.5Y is independently --H or --R.sup.5YA; [0063] --R.sup.5YA
is independently linear or branched saturated C.sub.1-6alkyl;
[0064] --R.sup.7 is independently --R.sup.7X or
--C(.dbd.O)R.sup.7X; [0065] each --R.sup.7X is independently:
[0066] --R.sup.7A, --R.sup.7B--R.sup.7C, --R.sup.7D, --R.sup.7E,
[0067] -L.sup.7-R.sup.7B, -L.sup.7-R.sup.7C, -L.sup.7-R.sup.7D, or
-L.sup.7-R.sup.7E; [0068] each -L.sup.7- is independently linear or
branched saturated C.sub.1-6alkylene; [0069] each --R.sup.7A is
independently linear or branched saturated C.sub.1-6alkyl, and is
optionally substituted with one or more substituents --W.sup.1;
[0070] each --R.sup.7B is saturated C.sub.3-6cycloalkyl, and is
optionally substituted with one or more substituents --W.sup.2;
[0071] each --R.sup.7D is non-aromatic C.sub.3-7heterocyclyl, and
is optionally substituted with one or more substituents --W.sup.2;
[0072] each --R.sup.7D is independently phenyl or naphthyl, and is
optionally substituted with one or more substituents --W.sup.3;
[0073] each --R.sup.7E is C.sub.6-12heteroaryl, and is optionally
substituted with one or more substituents --W.sup.3; [0074] each
--W.sup.1 is independently: [0075] --F, --Cl, --Br, --I,
--CF.sub.3, --OH, --OR.sup.W1, --OCF.sub.3, --NH.sub.2,
--NHR.sup.W1, --NR.sup.W1.sub.2, pyrrolidino, piperidino,
morpholino, piperazino, N--(R.sup.W1)-piperazino, --C(.dbd.O)OH,
--C(.dbd.O)OR.sup.W1, --C(.dbd.O)NH.sub.2, --C(.dbd.O)NHR.sup.W1,
--C(.dbd.O)NR.sup.W1.sub.2, --C(.dbd.O)-pyrrolidino,
--C(.dbd.O)-piperidino, --C(.dbd.O)-morpholino,
--C(.dbd.O)-piperazino, --C(.dbd.O)--N-(R.sup.W1)-piperazino,
--S(.dbd.O)R.sup.W1, --S(.dbd.O).sub.2.sup.RW1,
--S(.dbd.O).sub.2NH.sub.2, --S(.dbd.O).sub.2NHR.sup.W1,
--S(.dbd.O).sub.2NR.sup.W1.sub.2, --S(.dbd.O).sub.2pyrrolidino,
--S(.dbd.O).sub.2-piperidino, --S(.dbd.O).sub.2-morpholino,
--S(.dbd.O).sub.2-piperazino,
--S(.dbd.O).sub.2--N--(R.sup.W1)-piperazino, --CN, or --NO.sub.2;
[0076] wherein each --R.sup.W1 is independently linear or branched
saturated C.sub.1-6alkyl, phenyl, or --CH.sub.2-phenyl, wherein
each phenyl is optionally substituted with one or more groups
selected from --F, --Cl, --Br, --I, --R.sup.W11, --CF.sub.3, --OH,
--OR.sup.W11, and --OCF.sub.3, wherein each --R.sup.W11 is
independently linear or branched saturated C.sub.1-6alkyl; [0077]
each --W.sup.2 is independently: [0078] --F, --Cl, --Br, --I,
--R.sup.W2, --CF.sub.3, --OH, --OR.sup.W2, --OCF.sub.3, --NH.sub.2,
--NHR.sup.W2, --NR.sup.W2.sub.2, pyrrolidino, piperidino,
morpholino, piperazino, N--(R.sup.W2)-piperazino, --C(.dbd.O)OH,
--C(.dbd.O)OR.sup.W2, --C(.dbd.O)NH.sub.2, --C(.dbd.O)NHR.sup.W2,
--C(.dbd.O)NR.sup.W2.sub.2, --C(.dbd.O)-pyrrolidino,
--C(.dbd.O)-piperidino, --C(.dbd.O)-morpholino,
--C(.dbd.O)-piperazino, --C(.dbd.O)--N--(R.sup.W2)-piperazino,
--S(.dbd.O)R.sup.W2, --S(.dbd.O).sub.2R.sup.W2,
--S(.dbd.O).sub.2NH.sub.2, --S(.dbd.O).sub.2NHR.sup.W2,
--S(.dbd.O).sub.2NR.sup.W2.sub.2, --S(.dbd.O).sub.2pyrrolidino,
--S(.dbd.O).sub.2-piperidino, --S(.dbd.O).sub.2-morpholino,
--S(.dbd.O).sub.2-piperazino,
--S(.dbd.O).sub.2--N--(R.sup.W2)-piperazino, --CN, or --NO.sub.2;
[0079] wherein each --R.sup.W2 is independently linear or branched
saturated C.sub.1-6alkyl, phenyl, or --CH.sub.2-phenyl, wherein
each phenyl is optionally substituted with one or more groups
selected from --F, --Cl, --Br, --I, --R.sup.W22, --CF.sub.3, --OH,
--OR.sup.W22, and --OCF.sub.3, wherein each --R.sup.W22 is
independently linear or branched saturated C.sub.1-6alkyl; [0080]
each --W.sup.3 is independently: [0081] --F, --Cl, --Br, --I,
--R.sup.W3, --CF.sub.3, --OH, --OR.sup.W3, --OCF.sub.3, --NH.sub.2,
--NHR.sup.W3, --NR.sup.W3.sub.2, pyrrolidino, piperidino,
morpholino, piperazino, N--(R.sup.W3)-piperazino, --C(.dbd.O)OH,
--C(.dbd.O)OR.sup.W3, --C(.dbd.O)NH.sub.2, --C(.dbd.O)NHR.sup.W3,
--C(.dbd.O)NR.sup.W3.sub.2, --C(.dbd.O)-pyrrolidino,
--C(.dbd.O)-piperidino, --C(.dbd.O)-morpholino,
--C(.dbd.O)-piperazino, --C(.dbd.O)--N--(R.sup.W3)-piperazino,
--S(.dbd.O)R.sup.W3, --S(.dbd.O).sub.2R.sup.W3,
--S(.dbd.O).sub.2NH.sub.2, --S(.dbd.O).sub.2NHR.sup.W3,
--S(.dbd.O).sub.2NR.sup.W3.sub.2, --S(.dbd.O).sub.2pyrrolidino,
--S(.dbd.O).sub.2-piperidino, --S(.dbd.O).sub.2-morpholino,
--S(.dbd.O).sub.2-piperazino,
--S(.dbd.O).sub.2--N--(R.sup.W3)-piperazino, --CN, or --NO.sub.2;
[0082] wherein each --R.sup.W3 is independently linear or branched
saturated C.sub.1-6alkyl, phenyl, or --CH.sub.2-phenyl, wherein
each phenyl is optionally substituted with one or more groups
selected from --F, --Cl, --Br, --I, --R.sup.W33, --CF.sub.3, --OH,
--OR.sup.W33, and --OCF.sub.3, wherein each --R.sup.W33 is
independently linear or branched saturated C.sub.1-6alkyl; [0083]
--R.sup.3 is independently --R.sup.3A or --R.sup.3B; [0084]
--R.sup.3A is independently linear or branched saturated
C.sub.1-6alkyl; [0085] --R.sup.3B is saturated C.sub.3-7cycloalkyl;
[0086] --R.sup.2 is independently --H or --R.sup.2A; [0087]
--R.sup.2A is independently --F, --Cl, --Br, --I, --R.sup.2AA,
--CF.sub.3, --OH, --OR.sup.2AA, --OCF.sub.3, --NH.sub.2,
--NHR.sup.2AA, --NR.sup.2AA.sub.2, pyrrolidino, piperidino,
morpholino, piperazino, N--(R.sup.2AA)-piperazino, --SH,
--SR.sup.2AA, or --CN; [0088] each --R.sup.2AA is independently
linear or branched saturated C.sub.1-6alkyl; [0089] --R.sup.6 is
independently --H or --R.sup.6A; [0090] --R.sup.6A is independently
--F, --Cl, --Br, --I, --R.sup.6AA, --CF.sub.3, --OH, --OR.sup.6AA,
--OCF.sub.3, --NH.sub.2, --NHR.sup.6AA, --NR.sup.6AA.sub.2,
pyrrolidino, piperidino, morpholino, piperazino,
N--(R.sup.6AA)-piperazino, --SH, --SR.sup.6AA, or --CN; and [0091]
each --R.sup.6AA is independently linear or branched saturated
C.sub.1-6alkyl.
[0092] For the avoidance of doubt:
[0093] The index "C.sub.x-y" in terms such as
"C.sub.5-10heteroaryl", "C.sub.3-7heterocyclyl", and the like,
refers to the number of ring atoms, which may be carbon atoms or
heteroatoms (e.g., N, O, S). For example, pyridyl is an example of
a C.sub.6heteroaryl group, and piperidino is an example of a
C.sub.6heterocyclyl group.
[0094] The term "heteroaryl" refers to a group that is attached to
the rest of the molecule by an atom that is part of an aromatic
ring, wherein the aromatic ring is part of an aromatic ring system,
and the aromatic ring system has one or more heteroatoms (e.g., N,
O, S). For example, pyridyl is an example of a C.sub.6heteroaryl
group, and quinolyl is an example of a C.sub.10heteroaryl
group.
[0095] The term "heterocyclyl" refers to a group that is attached
to the rest of the molecule by a ring atom that is not part of an
aromatic ring (i.e., the ring is partially or fully saturated), and
the ring contains one or more heteroatoms (e.g., N, O, S). For
example, piperidino is an example of a C.sub.6heterocyclyl
group.
[0096] Unless otherwise indicated, where a compound is shown or
described which has one or more chiral centres, and two or more
stereoisomers are possible, all such stereoisomers are disclosed
and encompassed, both individually (e.g., as isolated from the
other stereoisomer(s)) and as mixtures (e.g., as equimolar or
non-equimolar mixtures of two or more stereoisomers). For example,
unless otherwise indicated, where a compound has one chiral centre,
each of the (R) and (S) enantiomers are disclosed and encompassed,
both individually (e.g., as isolated from the other enantiomer) and
as a mixture (e.g., as equimolar or non-equimolar mixtures of the
two enantiomers). For example, the initial carbon atom of a pendant
sec-butyl group, --CH(CH.sub.3)CH.sub.2CH.sub.3 is usually chiral,
and so gives rise to stereoisomers, e.g., (R) and (S) enantiomers
if it is the only chiral centre, each of which is disclosed and
encompassed.
[0097] The Group -L.sup.5X-
[0098] (2) A compound according to (1), wherein -L.sup.5X- is a
covalent single bond.
[0099] (3) A compound according to (1), wherein -L.sup.5X- is
-L.sup.5XA-.
[0100] The Group -L.sup.5XA-
[0101] (4) A compound according to any one of (1) to (3), wherein
-L.sup.5XA-, if present, is independently linear or branched
saturated C.sub.1-6alkylene.
[0102] (5) A compound according to any one of (1) to (3), wherein
-L.sup.5XA-, if present, is independently linear or branched
saturated C.sub.1-4alkylene, and is optionally substituted with one
or more groups selected from --OH and --OR.sup.L5X, wherein each
--R.sup.L5X is independently linear or branched saturated
C.sub.1-6alkyl or saturated C.sub.3-6cycloalkyl.
[0103] (6) A compound according to any one of (1) to (3), wherein
-L.sup.5XA-, if present, is independently linear or branched
saturated C.sub.1-4alkylene.
[0104] (7) A compound according to any one of (1) to (3), wherein
-L.sup.5XA-, if present, is independently --CH.sub.2--,
--CH(CH.sub.3)--, --C(CH.sub.3).sub.2--, --CH.sub.2CH.sub.2--,
--CH(CH.sub.3)CH.sub.2--, --CH.sub.2CH(CH.sub.3)--, or
--CH.sub.2CH.sub.2CH.sub.2--.
[0105] (8) A compound according to any one of (1) to (3), wherein
-L.sup.5XA-, if present, is independently --CH.sub.2--,
--CH(CH.sub.3)--, --C(CH.sub.3).sub.2--, or
--CH.sub.2CH.sub.2--.
[0106] (9) A compound according to any one of (1) to (3), wherein
-L.sup.5XA-, if present, is independently --CH.sub.2--,
--CH(CH.sub.3)--, or --C(CH.sub.3).sub.2--.
[0107] (10) A compound according to any one of (1) to (3), wherein
-L.sup.5XA, if present, is independently --CH.sub.2-- or
--CH.sub.2CH.sub.2--.
[0108] (11) A compound according to any one of (1) to (3), wherein
-L.sup.5XA-, if present, is --CH.sub.2--.
[0109] The Group --R.sup.L5X
[0110] (12) A compound according to any one of (1) to (11), wherein
each --R.sup.L5X, if present, is independently linear or branched
saturated C.sub.1-4alkyl or saturated C.sub.3-6cycloalkyl.
[0111] (13) A compound according to any one of (1) to (11), wherein
each --R.sup.L5X, if present, is independently linear or saturated
C.sub.1-4alkyl.
[0112] (14) A compound according to any one of (1) to (11), wherein
each --R.sup.L5X, if present, is independently -Me, -Et, -nPr,
-iPr, -nBu, -iBu, -sBu, or -tBu.
[0113] (15) A compound according to any one of (1) to (11), wherein
each --R.sup.L5X, if present, is independently -Me, -Et, -nPr, or
-iPr.
[0114] (16) A compound according to any one of (1) to (11), wherein
each --R.sup.L5X, if present, is independently -Me or -Et.
[0115] (17) A compound according to any one of (1) to (11), wherein
each --R.sup.L5X, if present, is -Me.
[0116] The Group -Q
[0117] (18) A compound according to any one of (1) to (17), wherein
-Q is a non-aromatic heterocyclic ring having from 5 to 7 ring
atoms, including at least one nitrogen ring atom, wherein the point
of attachment is via a ring carbon atom, and is substituted with
"n" groups -J, and is substituted with "m" groups --R.sup.Q.
[0118] (19) A compound according to any one of (1) to (17), wherein
-Q is a non-aromatic heterocyclic ring having from 5 to 7 ring
atoms, including at least one nitrogen ring atom, wherein the point
of attachment is via a ring nitrogen atom, and is substituted with
"n" groups -J, and is substituted with "m" groups --R.sup.Q.
[0119] (20) A compound according to any one of (1) to (17), wherein
-Q is pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl,
azepanyl, or diazepanyl, and is substituted with "n" groups -J, and
is substituted with "m" groups --R.sup.Q.
##STR00016##
[0120] (21) A compound according to any one of (1) to (17), wherein
-Q is pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl,
azepanyl, or diazepanyl, wherein the point of attachment is via a
ring carbon atom, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0121] (22) A compound according to any one of (1) to (17), wherein
-Q is pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl,
azepanyl, or diazepanyl, wherein the point of attachment is via a
ring nitrogen atom, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0122] (23) A compound according to any one of (1) to (17), wherein
-Q is pyrrolidinyl, piperidinyl, morpholinyl, or piperazinyl, and
is substituted with "n" groups -J, and is substituted with "m"
groups --R.sup.Q.
[0123] (24) A compound according to any one of (1) to (17), wherein
-Q is pyrrolidinyl, piperidinyl, morpholinyl, or piperazinyl,
wherein the point of attachment is via a ring carbon atom, and is
substituted with "n" groups -J, and is substituted with "m" groups
--R.sup.Q.
[0124] (25) A compound according to any one of (1) to (17), wherein
-Q is pyrrolidinyl, piperidinyl, morpholinyl, or piperazinyl,
wherein the point of attachment is via a ring nitrogen atom, and is
substituted with "n" groups -J, and is substituted with "m" groups
--R.sup.Q.
[0125] (26) A compound according to any one of (1) to (17), wherein
-Q is pyrrolidinyl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0126] (27) A compound according to any one of (1) to (17), wherein
-Q is pyrrolidinyl, wherein the point of attachment is via a ring
carbon atom, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0127] (28) A compound according to any one of (1) to (17), wherein
-Q is pyrrolidinyl, wherein the point of attachment is via a ring
nitrogen atom (i.e., pyrrolidino) and is substituted with "n"
groups -J, and is substituted with "m" groups --R.sup.Q.
[0128] (29) A compound according to any one of (1) to (17), wherein
-Q is pyrrolidin-2-yl, and is substituted with "n" groups -J, and
is substituted with "m" groups --R.sup.Q.
[0129] (30) A compound according to any one of (1) to (17), wherein
-Q is pyrrolidin-3-yl, and is substituted with "n" groups -J, and
is substituted with "m" groups --R.sup.Q.
[0130] (31) A compound according to any one of (1) to (17), wherein
-Q is piperidinyl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0131] (32) A compound according to any one of (1) to (17), wherein
-Q is piperidinyl, wherein the point of attachment is via a ring
carbon atom, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0132] (33) A compound according to any one of (1) to (17), wherein
-Q is piperidin-4-yl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0133] (34) A compound according to any one of (1) to (17), wherein
-Q is piperidin-3-yl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0134] (35) A compound according to any one of (1) to (17), wherein
-Q is piperidin-2-yl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0135] (36) A compound according to any one of (1) to (17), wherein
-Q is piperidinyl, wherein the point of attachment is via a ring
nitrogen atom (i.e., piperidino), and is substituted with "n"
groups -J, and is substituted with "m" groups --R.sup.Q.
[0136] (37) A compound according to any one of (1) to (17), wherein
-Q is morpholinyl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0137] (38) A compound according to any one of (1) to (17), wherein
-Q is morpholinyl, wherein the point of attachment is via a ring
carbon atom, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0138] (39) A compound according to any one of (1) to (17), wherein
-Q is morpholin-2-yl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0139] (40) A compound according to any one of (1) to (17), wherein
-Q is morpholin-3-yl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0140] (41) A compound according to any one of (1) to (17), wherein
-Q is morpholinyl, wherein the point of attachment is via a ring
nitrogen atom (i.e., morpholino), and is substituted with "n"
groups -J, and is substituted with "m" groups --R.sup.Q.
[0141] (42) A compound according to any one of (1) to (17), wherein
-Q is piperazinyl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0142] (43) A compound according to any one of (1) to (17), wherein
-Q is piperazinyl, wherein the point of attachment is via a ring
carbon atom, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0143] (44) A compound according to any one of (1) to (17), wherein
-Q is piperazin-2-yl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0144] (45) A compound according to any one of (1) to (17), wherein
-Q is piperazin-3-yl, and is substituted with "n" groups -J, and is
substituted with "m" groups --R.sup.Q.
[0145] (46) A compound according to any one of (1) to (17), wherein
-Q is piperazinyl, wherein the point of attachment is via a ring
nitrogen atom (i.e., piperazino), and is substituted with "n"
groups -J, and is substituted with "m" groups --R.sup.Q.
[0146] The Index "n"
[0147] (47) A compound according to any one of (1) to (46), wherein
"n" is 1 or 2.
[0148] (48) A compound according to any one of (1) to (46), wherein
"n" is 1.
[0149] (49) A compound according to any one of (1) to (46), wherein
"n" is 2.
[0150] (50) A compound according to any one of (1) to (46), wherein
"n" is 3.
[0151] The Index "m"
[0152] (51) A compound according to any one of (1) to (50), wherein
"m" is 0, 1 or 2.
[0153] (52) A compound according to any one of (1) to (50), wherein
"m" is 0 or 1.
[0154] (53) A compound according to any one of (1) to (50), wherein
"m" is 0.
[0155] (54) A compound according to any one of (1) to (50), wherein
"m" is 1.
[0156] (55) A compound according to any one of (1) to (50), wherein
"m" is 2.
[0157] (56) A compound according to any one of (1) to (50), wherein
"m" is 3.
[0158] Some Preferred Groups -Q
[0159] (57) A compound according to any one of (1) to (56), wherein
-Q is selected from:
##STR00017##
[0160] (58) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00018##
[0161] (59) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00019##
[0162] (60) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00020##
[0163] (61) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00021##
[0164] (62) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00022##
[0165] (63) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00023##
[0166] (64) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00024##
[0167] (65) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00025##
[0168] (66) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00026##
[0169] (67) A compound according to any one of (1) to (56), wherein
-Q is selected from:
##STR00027##
[0170] (68) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00028##
[0171] (69) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00029##
[0172] (70) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00030##
[0173] (71) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00031##
[0174] (72) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00032##
[0175] (73) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00033##
[0176] (74) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00034##
[0177] (75) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00035##
[0178] (76) A compound according to any one of (1) to (56), wherein
-Q is selected from:
##STR00036##
[0179] (77) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00037##
[0180] (78) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00038##
[0181] (79) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00039##
[0182] (80) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00040##
[0183] (81) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00041##
[0184] (82) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00042##
[0185] (83) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00043##
[0186] (84) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00044##
[0187] (85) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00045##
[0188] (86) A compound according to any one of (1) to (56), wherein
-Q is selected from:
##STR00046##
[0189] (87) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00047##
[0190] (88) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00048##
[0191] (89) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00049##
[0192] (90) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00050##
[0193] (91) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00051##
[0194] (92) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00052##
[0195] (93) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00053##
[0196] (94) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00054##
[0197] (95) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00055##
[0198] (96) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00056##
[0199] (97) A compound according to any one of (1) to (56), wherein
-Q is:
##STR00057##
[0200] The Group -J
[0201] (98) A compound according to any one of (1) to (97), wherein
each -J is independently --OH or --OR.sup.J.
[0202] (99) A compound according to any one of (1) to (97), wherein
each -J is independently --OH or -L.sup.J-OH.
[0203] (100) A compound according to any one of (1) to (97),
wherein each -J is independently -L.sup.J-OH or
-L.sup.J-OR.sup.J.
[0204] (101) A compound according to any one of (1) to (97),
wherein each -J is --OH.
[0205] (102) A compound according to any one of (1) to (97),
wherein each -J is --OR.sup.J.
[0206] (103) A compound according to any one of (1) to (97),
wherein each -J is -L.sup.J-OH.
[0207] (104) A compound according to any one of (1) to (97),
wherein each -J is -L.sup.J-OR.sup.J.
[0208] The Group --R.sup.J
[0209] (105) A compound according to any one of (1) to (104),
wherein each --R.sup.J, if present, is independently linear or
branched saturated C.sub.1-4alkyl or saturated
C.sub.3-6cycloalkyl.
[0210] (106) A compound according to any one of (1) to (104),
wherein each --R.sup.J, if present, is independently linear or
branched saturated C.sub.1-4alkyl.
[0211] (107) A compound according to any one of (1) to (104),
wherein each --R.sup.J, if present, is independently -Me, -Et,
-nPr, -iPr, -nBu, -iBu, -sBu, or -tBu.
[0212] (108) A compound according to any one of (1) to (104),
wherein each --R.sup.J, if present, is independently -Me, -Et,
-nPr, or -iPr.
[0213] (109) A compound according to any one of (1) to (104),
wherein each --R.sup.J, if present, is independently -Me or
-Et.
[0214] (110) A compound according to any one of (1) to (104),
wherein each --R.sup.J, if present, is -Me.
[0215] The Group -L.sup.J-
[0216] (111) A compound according to any one of (1) to (110),
wherein each -L.sup.J-, if present, is independently linear or
branched saturated C.sub.1-4alkylene.
[0217] (112) A compound according to any one of (1) to (110),
wherein each -L.sup.J-, if present, is independently --CH.sub.2--,
--CH(CH.sub.3)--, --C(CH.sub.3).sub.2--, --CH.sub.2CH.sub.2--,
--CH(CH.sub.3)CH.sub.2--, --CH.sub.2CH(CH.sub.3)--, or
--CH.sub.2CH.sub.2CH.sub.2--.
[0218] (113) A compound according to any one of (1) to (110),
wherein each -L.sup.J-, if present, is independently --CH.sub.2--,
--CH(CH.sub.3)--, --C(CH.sub.3).sub.2--, or
--CH.sub.2CH.sub.2--.
[0219] (114) A compound according to any one of (1) to (110),
wherein each -L.sup.J-, if present, is independently --CH.sub.2--,
--CH(CH.sub.3)--, or --C(CH.sub.3).sub.2--.
[0220] (115) A compound according to any one of (1) to (110),
wherein each -L.sup.J-, if present, is independently --CH.sub.2--
or --CH.sub.2CH.sub.2--.
[0221] (116) A compound according to any one of (1) to (110),
wherein each -L.sup.J-, if present, is independently
--CH.sub.2--.
[0222] The Group --R.sup.Q
[0223] (117) A compound according to any one of (1) to (116),
wherein each --R.sup.Q, if present, is independently --F, --Cl,
--Br, --I, --R.sup.QA, --CF.sub.3, --OH, --OR.sup.QA, --OCF.sub.3,
--NH.sub.2, --NHR.sup.QA, --NR.sup.QA.sub.2, pyrrolidino,
piperidino, morpholino, piperazino, or
N--(R.sup.QA)-piperazino.
[0224] (118) A compound according to any one of (1) to (116),
wherein each --R.sup.Q, if present, is independently --F, --Cl,
--Br, --I, --R.sup.QA, --OH, or --OR.sup.QA.
[0225] (119) A compound according to any one of (1) to (116),
wherein each --R.sup.Q, if present, is independently --F, --Cl,
--Br, or --I.
[0226] The Group --R.sup.QA
[0227] (120) A compound according to any one of (1) to (119),
wherein each --R.sup.QA, if present, is independently linear or
branched saturated C.sub.1-4alkyl or saturated
C.sub.3-6cycloalkyl.
[0228] (121) A compound according to any one of (1) to (119),
wherein each --R.sup.QA, if present, is independently linear or
branched C.sub.1-4alkyl.
[0229] (122) A compound according to any one of (1) to (119),
wherein each --R.sup.QA, if present, is independently -Me, -Et,
-nPr, -iPr, -nBu, -iBu, -sBu, or -tBu.
[0230] (123) A compound according to any one of (1) to (119),
wherein each --R.sup.QA, if present, is independently -Me, -Et,
-nPr, or -iPr.
[0231] (124) A compound according to any one of (1) to (119),
wherein each --R.sup.QA, if present, is independently -Me or
-Et.
[0232] (125) A compound according to any one of (1) to (119),
wherein each --R.sup.QA, if present, is independently -Me.
[0233] The Group --R.sup.5Y
[0234] (126) A compound according to any one of (1) to (125),
wherein --R.sup.5Y is --H.
[0235] (127) A compound according to any one of (1) to (125),
wherein --R.sup.5Y is --R.sup.5YA.
[0236] The Group --R.sup.5YA
[0237] (128) A compound according to any one of (1) to (127),
wherein --R.sup.5YA, if present, is independently linear or
branched saturated C.sub.1-4alkyl.
[0238] (129) A compound according to any one of (1) to (127),
wherein --R.sup.5YA, if present, is independently -Me, -Et, -nPr,
-iPr, -nBu, -iBu, -sBu, or -tBu.
[0239] (130) A compound according to any one of (1) to (127),
wherein --R.sup.5YA, if present, is independently -Me, -Et, -nPr,
or -iPr.
[0240] (131) A compound according to any one of (1) to (127),
wherein --R.sup.5YA, if present, is independently -Me or -Et.
[0241] (132) A compound according to any one of (1) to (127),
wherein --R.sup.5YA, if present, is independently -Me.
[0242] The Group --R.sup.7
[0243] (133) A compound according to any one of (1) to (132),
wherein --R.sup.7 is --R.sup.7X.
[0244] (134) A compound according to any one of (1) to (132),
wherein --R.sup.7 is --C(.dbd.O)R.sup.7X.
[0245] The Group --R.sup.7X
[0246] (135) A compound according to any one of (1) to (134),
wherein each --R.sup.7X is independently: [0247] --R.sup.7B,
--R.sup.7C, --R.sup.7D, --R.sup.7E, [0248] -L.sup.7-R.sup.7B,
-L.sup.7-R.sup.7C, -L.sup.7-R.sup.7D, or -L.sup.7-R.sup.7E.
[0249] (136) A compound according to any one of (1) to (134),
wherein each --R.sup.7X is independently: [0250] --R.sup.7C,
--R.sup.7D, --R.sup.7E, [0251] -L.sup.7R.sup.7B, -L.sup.7-R.sup.7D,
or -L.sup.7-R.sup.7E.
[0252] (137) A compound according to any one of (1) to (134),
wherein each --R.sup.7X is independently: [0253] -L.sup.7-R.sup.7B,
-L.sup.7-R.sup.7D, or -L.sup.7-R.sup.7E.
[0254] (138) A compound according to any one of (1) to (134),
wherein each --R.sup.7X is -L.sup.7-R.sup.7D.
[0255] (139) A compound according to any one of (1) to (134),
wherein each --R.sup.7X is --R.sup.7A.
[0256] (140) A compound according to any one of (1) to (134),
wherein each --R.sup.7X is --R.sup.7B.
[0257] (141) A compound according to any one of (1) to (134),
wherein each --R.sup.7X is --R.sup.7C.
[0258] (142) A compound according to any one of (1) to (134),
wherein each --R.sup.7X is --R.sup.7D.
[0259] (143) A compound according to any one of (1) to (134),
wherein each --R.sup.7X is --R.sup.7E.
[0260] (144) A compound according to any one of (1) to (134),
wherein each --R.sup.7X is -L.sup.7--R.sup.7B.
[0261] (145) A compound according to any one of (1) to (134),
wherein each --R.sup.7X is -L.sup.7-R.sup.7C.
[0262] (146) A compound according to any one of (1) to (134),
wherein each --R.sup.7X is -L.sup.7--R.sup.7D.
[0263] (147) A compound according to any one of (1) to (134),
wherein each --R.sup.7X is -L.sup.7-R.sup.7E.
[0264] The Group -L.sup.7-
[0265] (148) A compound according to any one of (1) to (147),
wherein each -L.sup.7-, if present, is independently linear or
branched saturated C.sub.1-4alkylene.
[0266] (149) A compound according to any one of (1) to (147),
wherein each -L.sup.7-, if present, is independently --CH.sub.2--,
--CH(CH.sub.3)--, --C(CH.sub.3).sub.2--, --CH.sub.2CH.sub.2--,
--CH(CH.sub.3)CH.sub.2--, --CH.sub.2CH(CH.sub.3)--, or
--CH.sub.2CH.sub.2CH.sub.2--.
[0267] (150) A compound according to any one of (1) to (147),
wherein each -L.sup.7-, if present, is independently --CH.sub.2--,
--CH(CH.sub.3)--, --C(CH.sub.3).sub.2--, or
--CH.sub.2CH.sub.2--.
[0268] (151) A compound according to any one of (1) to (147),
wherein each -L.sup.7-, if present, is independently --CH.sub.2--,
--CH(CH.sub.3)--, or --C(CH.sub.3).sub.2--.
[0269] (152) A compound according to any one of (1) to (147),
wherein each -L.sup.7-, if present, is independently --CH2-- or
--CH.sub.2CH.sub.2--.
[0270] (153) A compound according to any one of (1) to (147),
wherein each -L.sup.7-, if present, is --CH.sub.2--.
[0271] The Group --R.sup.7A
[0272] (154) A compound according to any one of (1) to (153),
wherein each --R.sup.7A, if present, is independently linear or
branched saturated C.sub.1-6alkyl.
[0273] (155) A compound according to any one of (1) to (153),
wherein each --R.sup.7A, if present, is independently linear or
branched saturated C.sub.1-4 alkyl, and is optionally substituted
with one or more substituents --W.sup.1.
[0274] (156) A compound according to any one of (1) to (153),
wherein each --R.sup.7A, if present, is independently linear or
branched saturated C.sub.1-4alkyl.
[0275] (157) A compound according to any one of (1) to (153),
wherein each --R.sup.7A, if present, is independently -Me, -Et,
-nPr, -iPr, -nBu, -iBu, -sBu, or -tBu, and is optionally
substituted with one or more substituents --W.sup.1.
[0276] (158) A compound according to any one of (1) to (153),
wherein each --R.sup.7A, if present, is independently -Me, -Et,
-nPr, -iPr, -nBu, -iBu, -sBu, or -tBu.
[0277] (159) A compound according to any one of (1) to (153),
wherein each --R.sup.7A, if present, is independently -Me, -Et,
-nPr, or -iPr, and is optionally substituted with one or more
substituents --W.sup.1.
[0278] (160) A compound according to any one of (1) to (153),
wherein each --R.sup.7A, if present, is independently -Me, -Et,
-nPr, or -iPr.
[0279] (161) A compound according to any one of (1) to (153),
wherein each --R.sup.7A, if present, is independently -Me or -Et,
and is optionally substituted with one or more substituents
--W.sup.1.
[0280] (162) A compound according to any one of (1) to (153),
wherein each --R.sup.7A, if present, is independently -Me or
-Et.
[0281] (163) A compound according to any one of (1) to (153),
wherein each --R.sup.7A, if present, is -Me, and is optionally
substituted with one or more substituents --W.sup.1.
[0282] (164) A compound according to any one of (1) to (153),
wherein each --R.sup.7A, if present, is -Me.
[0283] The Group --R.sup.7B
[0284] (165) A compound according to any one of (1) to (164),
wherein each --R.sup.7B, if present, is independently cyclopropyl,
cyclobutyl, cyclopentyl, or cyclohexyl, and is optionally
substituted with one or more substituents --W.sup.2.
[0285] (166) A compound according to any one of (1) to (164),
wherein each --R.sup.7B, if present, is independently cyclopropyl,
cyclobutyl, cyclopentyl, or cyclohexyl.
[0286] (167) A compound according to any one of (1) to (164),
wherein each --R.sup.7B, if present, is independently cyclopentyl
or cyclohexyl, and is optionally substituted with one or more
substituents --W.sup.2.
[0287] (168) A compound according to any one of (1) to (164),
wherein each --R.sup.7B, if present, is independently cyclopentyl
or cyclohexyl.
[0288] (169) A compound according to any one of (1) to (164),
wherein each --R.sup.7B, if present, is cyclohexyl, and is
optionally substituted with one or more substituents --W.sup.2.
[0289] (170) A compound according to any one of (1) to (164),
wherein each --R.sup.7B, if present, is cyclohexyl.
[0290] The Group --R.sup.7C
[0291] (171) A compound according to any one of (1) to (170),
wherein each --R.sup.7C, if present, is independently pyrrolidinyl,
piperidinyl, morpholinyl, piperazinyl, tetrahydrofuranyl,
tetrahydropyranyl, dixoanyl, azepanyl, or diazepanyl, and is
optionally substituted with one or more substituents --W.sup.2.
[0292] (172) A compound according to any one of (1) to (170),
wherein each --R.sup.7C, if present, is independently pyrrolidinyl,
piperidinyl, morpholinyl, piperazinyl, tetrahydrofuranyl,
tetrahydropyranyl, or dixoanyl, and is optionally substituted with
one or more substituents --W.sup.2.
[0293] (173) A compound according to any one of (1) to (170),
wherein each --R.sup.7C, if present, is independently pyrrolidinyl,
piperidinyl, morpholinyl, or piperazinyl, and is optionally
substituted with one or more substituents --W.sup.2.
[0294] (174) A compound according to any one of (1) to (170),
wherein each --R.sup.7C, if present, is independently piperidinyl,
and is optionally substituted with one or more substituents
--W.sup.2.
[0295] The Group --R.sup.7D
[0296] (175) A compound according to any one of (1) to (174),
wherein each --R.sup.7D, if present, is phenyl, and is optionally
substituted with one or more substituents --.sup.3.
[0297] (176) A compound according to any one of (1) to (174),
wherein each --R.sup.7D, if present, is phenyl.
[0298] (177) A compound according to any one of (1) to (174),
wherein each --R.sup.7D, if present, is naphthyl, and is optionally
substituted with one or more substituents --W.sup.3.
[0299] (178) A compound according to any one of (1) to (174),
wherein each --R.sup.7D, if present, is naphthyl.
[0300] The Group --R.sup.7E
[0301] (179) A compound according to any one of (1) to (178),
wherein each --R.sup.7E, if present, is C.sub.5-10heteroaryl, and
is optionally substituted with one or more substituents
--W.sup.3.
[0302] (180) A compound according to any one of (1) to (178),
wherein each --R.sup.7E, if present, is C.sub.5-10heteroaryl.
[0303] (181) A compound according to any one of (1) to (178),
wherein each --R.sup.7E, if present, is C.sub.5-6heteroaryl, and is
optionally substituted with one or more substituents --W.sup.3.
[0304] (182) A compound according to any one of (1) to (178),
wherein each --R.sup.7E, if present, is C.sub.5-6heteroaryl.
[0305] (183) A compound according to any one of (1) to (178),
wherein each --R.sup.7E, if present, is C.sub.9-10heteroaryl, and
is optionally substituted, for example, with one or more
substituents --W.sup.3.
[0306] (184) A compound according to any one of (1) to (178),
wherein each --R.sup.7E, if present, is C.sub.9-10heteroaryl.
[0307] (185) A compound according to any one of (1) to (178),
wherein each --R.sup.7E, if present, is independently furanyl,
thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl,
thiazolyl, isothiazolyl, triazolyl (e.g., 1H-[1,2,3]triazolyl,
2H-[1,2,3]triazolyl, 4H-[1,2,4]triazolyl, 1H-[1,2,4]triazolyl),
oxadiazolyl (e.g., [1,2,3]oxadiazolyl, furazanyl,
[1,3,4]oxadiazolyl, [1,2,4]oxadiazolyl), thiadiazolyl (e.g.,
[1,2,3]thiadiazolyl, [1,2,5]thiadiazolyl, [1,3,4]thiadiazolyl,
[1,2,4]thiadiazolyl), tetrazolyl, pyridyl, pyrimidinyl, pyrazinyl,
pyridazinyl, or triazinyl (e.g., [1,3,5]-triazinyl), and is
optionally substituted with one or more substituents --W.sup.3.
[0308] (186) A compound according to any one of (1) to (178),
wherein each --R.sup.7E, if present, is independently furanyl,
thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl,
thiazolyl, isothiazolyl, pyridyl, pyrimidinyl, pyrazinyl, or
pyridazinyl, and is optionally substituted, for example, with one
or more substituents --W.sup.3.
[0309] (187) A compound according to any one of (1) to (178),
wherein each --R.sup.7E, if present, is independently furanyl,
thienyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl,
thiazolyl, or isothiazolyl, and is optionally substituted, for
example, with one or more substituents --W.sup.3.
[0310] (188) A compound according to any one of (1) to (178),
wherein each --R.sup.7E, if present, is independently pyridyl,
pyrimidinyl, pyrazinyl, or pyridazinyl, and is optionally
substituted, for example, with one or more substituents
--W.sup.3.
[0311] (189) A compound according to any one of (1) to (178),
wherein each --R.sup.7E, if present, is independently indolyl,
indazolyl, benzofuranyl, benzothienyl, benzimidazolyl,
benzothiazolyl, benzoxazolyl, benzoisoxazolyl, quinolinyl,
isoquinolinyl, cinnolinyl, quinazolinyl, phthalazinyl, or
quinoxalinyl, and is optionally substituted, for example, with one
or more substituents --W.sup.3.
[0312] (190) A compound according to any one of (1) to (178),
wherein each --R.sup.7E, if present, is benzothiazolyl, and is
optionally substituted, for example, with one or more substituents
--W.sup.3.
[0313] The Group --W.sup.1
[0314] (191) A compound according to any one of (1) to (190),
wherein each --W.sup.1, if present, is independently: --F, --Cl,
--Br, --I, --CF.sub.3, --OH, --OR.sup.W1, --OCF.sub.3, --NH.sub.2,
--NHR.sup.W1, --NR.sup.W1.sub.2, pyrrolidino, piperidino,
morpholino, piperazino, or N--(R.sup.W1)-piperazino.
[0315] (192) A compound according to any one of (1) to (190),
wherein each --W.sup.1, if present, is independently: --F, --Cl,
--Br, --I, --CF.sub.3, --OH, --OR.sup.W1, or --OCF.sub.3.
[0316] The Group --R.sup.W1
[0317] (193) A compound according to any one of (1) to (192),
wherein each --R.sup.W1, if present, is independently linear or
branched saturated C.sub.1-4alkyl, phenyl, or --CH.sub.2-phenyl,
wherein each phenyl is optionally substituted with one or more
groups selected from --F, --Cl, --Br, --I, --R.sup.W11, --CF.sub.3,
--OH, --OR.sup.W11, and --OCF.sub.3, wherein each --R.sup.W11 is
independently linear or branched saturated C.sub.1-4alkyl.
[0318] (194) A compound according to any one of (1) to (192),
wherein each --R.sup.W1, if present, is independently linear or
branched saturated C.sub.1-4alkyl, phenyl, or
--CH.sub.2-phenyl.
[0319] (195) A compound according to any one of (1) to (192),
wherein each --R.sup.W1, if present, is independently linear or
branched saturated C.sub.1-4alkyl.
[0320] (196) A compound according to any one of (1) to (192),
wherein each --R.sup.W1, if present, is independently -Me, -Et,
-nPr, -iPr, -nBu, -iBu, -sBu, or -tBu.
[0321] (197) A compound according to any one of (1) to (192),
wherein each --R.sup.W1, if present, is independently -Me, -Et,
-nPr, or -iPr.
[0322] (198) A compound according to any one of (1) to (192),
wherein each --R.sup.W1, if present, is independently -Me or
-Et.
[0323] (199) A compound according to any one of (1) to (192),
wherein each --R.sup.W1, if present, is -Me.
[0324] The Group --R.sup.W11
[0325] (200) A compound according to any one of (1) to (199),
wherein each --R.sup.W11, if present, is independently linear or
branched saturated C.sub.1-4alkyl.
[0326] (201) A compound according to any one of (1) to (199),
wherein each --R.sup.W11, if present, is independently -Me, -Et,
-nPr, -iPr, -nBu, -iBu, -sBu, or -tBu.
[0327] (202) A compound according to any one of (1) to (199),
wherein each --R.sup.W11, if present, is independently -Me, -Et,
-nPr, or -iPr.
[0328] (203) A compound according to any one of (1) to (199),
wherein each --R.sup.W11, if present, is independently -Me or
-Et.
[0329] (204) A compound according to any one of (1) to (199),
wherein each --R.sup.W11, if present, is -Me.
[0330] The Group --W.sup.2
[0331] (205) A compound according to any one of (1) to (204),
wherein each --W.sup.2, if present, is independently: --F, --Cl,
--Br, --I, --CF.sub.3, --OH, --OR.sup.W2, --OCF.sub.3, --NH.sub.2,
--NHR.sup.W2, --NR.sup.W2.sub.2, pyrrolidino, piperidino,
morpholino, piperazino, or N--(R.sup.W2)-piperazino.
[0332] (206) A compound according to any one of (1) to (204),
wherein each --W.sup.2, if present, is independently: --F, --Cl,
--Br, --I, --CF.sub.3, --OH, --OR.sup.W2, or --OCF.sub.3.
[0333] The Group --R.sup.W2
[0334] (207) A compound according to any one of (1) to (206),
wherein each --R.sup.W2, if present, is independently linear or
branched saturated C.sub.1-4alkyl, phenyl, or --CH.sub.2-phenyl,
wherein each phenyl is optionally substituted with one or more
groups selected from --F, --Cl, --Br, --I, --R.sup.W22, --CF.sub.3,
--OH, --OR.sup.W22, and --OCF.sub.3, wherein each --R.sup.W22 is
independently linear or branched saturated C.sub.1-4alkyl.
[0335] (208) A compound according to any one of (1) to (206),
wherein each --R.sup.W2, if present, is independently linear or
branched saturated C.sub.1-4alkyl, phenyl, or
--CH.sub.2-phenyl.
[0336] (209) A compound according to any one of (1) to (206),
wherein each --R.sup.W2, if present, is independently linear or
branched saturated C.sub.1-4alkyl.
[0337] (210) A compound according to any one of (1) to (206),
wherein each --R.sup.W2, if present, is independently -Me, -Et,
-nPr, -iPr, -nBu, -iBu, -sBu, or -tBu.
[0338] (211) A compound according to any one of (1) to (206),
wherein each --R.sup.W2, if present, is independently -Me, -Et,
-nPr, or -iPr.
[0339] (212) A compound according to any one of (1) to (206),
wherein each --R.sup.W2, if present, is independently -Me or
-Et.
[0340] (213) A compound according to any one of (1) to (206),
wherein each --R.sup.W2, if present, is -Me.
[0341] The Group --R.sup.W22
[0342] (214) A compound according to any one of (1) to (213),
wherein each --R.sup.W22, if present, is independently linear or
branched saturated C.sub.1-4alkyl.
[0343] (215) A compound according to any one of (1) to (213),
wherein each --R.sup.W22, if present, is independently -Me, -Et,
-nPr, -iPr, -nBu, -iBu, -sBu, or -tBu.
[0344] (216) A compound according to any one of (1) to (213),
wherein each --R.sup.W22, if present, is independently -Me, -Et,
-nPr, or -iPr.
[0345] (217) A compound according to any one of (1) to (213),
wherein each --R.sup.W22, if present, is independently -Me or
-Et.
[0346] (218) A compound according to any one of (1) to (213),
wherein each --R.sup.W22, if present, is -Me.
[0347] The Group --W.sup.3
[0348] (219) A compound according to any one of (1) to (218),
wherein each --W.sup.3, if present, is independently: --F, --Cl,
--Br, --I, --CF.sub.3, --OH, --OR.sup.W3, --OCF.sub.3, --NH.sub.2,
--NHR.sup.W3, --NR.sup.W3.sub.2, pyrrolidino, piperidino,
morpholino, piperazino, or N--(R.sup.W3)-piperazino.
[0349] (220) A compound according to any one of (1) to (218),
wherein each --W.sup.3, if present, is independently: --F, --Cl,
--Br, --I, --CF.sub.3, --OH, --OR.sup.W3, or --OCF.sub.3.
[0350] The Group --R.sup.W3
[0351] (221) A compound according to any one of (1) to (220),
wherein each --R.sup.W3, if present, is independently linear or
branched saturated C.sub.1-4alkyl, phenyl, or --CH.sub.2-phenyl,
wherein each phenyl is optionally substituted with one or more
groups selected from --F, --Cl, --Br, --I, --R.sup.W33, --CF.sub.3,
--OH, --OR.sup.W33, and --OCF.sub.3, wherein each --R.sup.W33 is
independently linear or branched saturated C.sub.1-4alkyl.
[0352] (222) A compound according to any one of (1) to (220),
wherein each --R.sup.W3, if present, is independently linear or
branched saturated C.sub.1-4alkyl, phenyl, or
--CH.sub.2-phenyl.
[0353] (223) A compound according to any one of (1) to (220),
wherein each --R.sup.W3, if present, is independently linear or
branched saturated C.sub.1-4alkyl.
[0354] (224) A compound according to any one of (1) to (220),
wherein each --R.sup.W3, if present, is independently linear or
branched saturated C.sub.1-4alkyl.
[0355] (225) A compound according to any one of (1) to (220),
wherein each --R.sup.W3, if present, is -Me.
[0356] The Group --R.sup.W33
[0357] (226) A compound according to any one of (1) to (225),
wherein each --R.sup.W33, if present, is independently linear or
branched saturated C.sub.1-4alkyl.
[0358] (227) A compound according to any one of (1) to (225),
wherein each --R.sup.W33, if present, is independently -Me, -Et,
-nPr, -iPr, -nBu, -iBu, -sBu, or -tBu.
[0359] (228) A compound according to any one of (1) to (225),
wherein each --R.sup.W33, if present, is independently -Me, -Et,
-nPr, or -iPr.
[0360] (229) A compound according to any one of (1) to (225),
wherein each --R.sup.W33, if present, is independently -Me or
-Et.
[0361] (230) A compound according to any one of (1) to (225),
wherein each --R.sup.W33, if present, is -Me.
[0362] The Group --R.sup.3
[0363] (231) A compound according to any one of (1) to (230),
wherein --R.sup.3 is --R.sup.3A.
[0364] (232) A compound according to any one of (1) to (230),
wherein --R.sup.3 is --R.sup.3B.
[0365] The Group --R.sup.3A
[0366] (233) A compound according to any one of (1) to (232),
wherein --R.sup.3A, if present, is independently -Me, -Et, -nPr,
-iPr, -nBu, -iBu, -sBu, or -tBu, n-pentyl, t-pentyl, neo-pentyl,
iso-pentyl, sec-pentyl, 3-pentyl, 1-hexyl, 2-hexyl, 3-hexyl,
3-methyl-1-pentyl, 4-methyl-1-pentyl, 4-methyl-2-pentyl,
4-methyl-3-pentyl, 2-methyl-2-pentyl, 2-methyl-1-pentyl,
2-methyl-2-pentyl, 3,3-dimethyl-1-butyl, 3,3-dimethyl-2-butyl,
3-methyl-1-pentyl, 3-methyl-2-pentyl, 3-methyl-3-pentyl,
2,2-dimethyl-1-butyl, 2,3-dimethyl-1-butyl, or
2,3-dimethyl-2-butyl.
[0367] (234) A compound according to any one of (1) to (232),
wherein --R.sup.3A, if present, is independently linear or branched
saturated C.sub.1-4alkyl.
[0368] (235) A compound according to any one of (1) to (232),
wherein --R.sup.3A, if present, is independently -Me, -Et, -nPr,
-iPr, -nBu, -iBu, -sBu, or -tBu.
[0369] (236) A compound according to any one of (1) to (232),
wherein --R.sup.3A, if present, is independently -Me, -Et, -nPr, or
-iPr.
[0370] (237) A compound according to any one of (1) to (232),
wherein --R.sup.3A, if present, is -iPr.
[0371] The Group --R.sup.3B
[0372] (238) A compound according to any one of (1) to (237),
wherein --R.sup.3B, if present, is independently cyclopropyl,
cyclobutyl, cyclopentyl, or cyclohexyl.
[0373] (239) A compound according to any one of (1) to (237),
wherein --R.sup.3B, if present, is independently cyclopropyl or
cyclobutyl.
[0374] (240) A compound according to any one of (1) to (237),
wherein --R.sup.3B, if present, is cyclopropyl.
[0375] (241) A compound according to any one of (1) to (237),
wherein --R.sup.3B, if present, is cyclobutyl.
[0376] The Group --R.sup.2
[0377] (242) A compound according to any one of (1) to (241),
wherein --R.sup.2 is --H.
[0378] (243) A compound according to any one of (1) to (241),
wherein --R.sup.2 is --R.sup.2A.
[0379] The Group --R.sup.2A
[0380] (244) A compound according to any one of (1) to (243),
wherein --R.sup.2A, if present, is independently --F, --Cl, --Br,
--I, --R.sup.2AA, --CF.sub.3, --OH, --OR.sup.2AA, --OCF.sub.3,
--NH.sub.2, --NHR.sup.2AA, --NR.sup.2AA.sub.2, pyrrolidino,
piperidino, morpholino, piperazino, or
N--(R.sup.2AA)-piperazino.
[0381] (245) A compound according to any one of (1) to (243),
wherein --R.sup.2A, if present, is independently --F, --Cl, --Br,
--I, --R.sup.2AA, --OH, or --OR.sup.2AA.
[0382] (246) A compound according to any one of (1) to (243),
wherein --R.sup.2A, if present, is independently --F, --Cl, --Br,
or --I.
[0383] The Group --R.sup.2AA
[0384] (247) A compound according to any one of (1) to (246),
wherein each --R.sup.2AA, if present, is independently linear or
branched saturated C.sub.1-4alkyl.
[0385] (248) A compound according to any one of (1) to (246),
wherein each --R.sup.2AA, if present, is independently -Me, -Et,
-nPr, -iPr, -nBu, -iBu, -sBu, or -tBu.
[0386] (249) A compound according to any one of (1) to (246),
wherein each --R.sup.2AA, if present, is independently -Me, -Et,
-nPr, or -iPr.
[0387] (250) A compound according to any one of (1) to (246),
wherein each --R.sup.2AA, if present, is independently -Me or
-Et.
[0388] (251) A compound according to any one of (1) to (246),
wherein each --R.sup.2AA, if present, is -Me.
[0389] The Group --R.sup.6
[0390] (252) A compound according to any one of (1) to (251),
wherein --R.sup.6 is --H.
[0391] (253) A compound according to any one of (1) to (251),
wherein --R.sup.6 is --R.sup.6A.
[0392] The Group --R.sup.6A
[0393] (254) A compound according to any one of (1) to (253),
wherein --R.sup.6A, if present, is independently --F, --Cl, --Br,
--I, --R.sup.6AA, --CF.sub.3, --OH, --OR.sup.6AA, --OCF.sub.3,
--NH.sub.2, --NHR.sup.6AA, --NR.sup.6AA.sub.2, pyrrolidino,
piperidino, morpholino, piperazino, or
N--(R.sup.6AA)-piperazino.
[0394] (255) A compound according to any one of (1) to (253),
wherein --R.sup.6A, if present, is independently --F, --Cl, --Br,
--I, --R.sup.6AA, --OH, or --OR.sup.6AA.
[0395] (256) A compound according to any one of (1) to (253),
wherein --R.sup.6A, if present, is independently --F, --Cl, --Br,
or --I.
[0396] The Group --R.sup.6AA
[0397] (257) A compound according to any one of (1) to (256),
wherein each --R.sup.6AA, if present, is independently linear or
branched saturated C.sub.1-4alkyl.
[0398] (258) A compound according to any one of (1) to (256),
wherein each --R.sup.6AA, if present, is independently -Me, -Et,
-nPr, -iPr, -nBu, -iBu, -sBu, or -tBu.
[0399] (259) A compound according to any one of (1) to (256),
wherein each --R.sup.6AA, if present, is independently -Me, -Et,
-nPr, or -iPr.
[0400] (260) A compound according to any one of (1) to (256),
wherein each --R.sup.6AA, if present, is independently -Me or
-Et.
[0401] (261) A compound according to any one of (1) to (256),
wherein each --R.sup.6AA, if present, is -Me.
[0402] Specific Compounds
[0403] (262) A compound according to (1), selected from compounds
of the following formulae and pharmaceutically acceptable salts,
hydrates, and solvates thereof:
TABLE-US-00002 Compound No. Structure PPDA-001 ##STR00058##
PPDA-002 ##STR00059## PPDA-003 ##STR00060## PPDA-004 ##STR00061##
PPDA-005 ##STR00062## PPDA-006 ##STR00063## PPDA-007 ##STR00064##
PPDA-008 ##STR00065## PPDA-009 ##STR00066## PPDA-010 ##STR00067##
PPDA-011 ##STR00068## PPDA-012 ##STR00069## PPDA-013 ##STR00070##
PPDA-014 ##STR00071## PPDA-015 ##STR00072## PPDA-016 ##STR00073##
PPDA-017 ##STR00074## PPDA-018 ##STR00075## PPDA-019 ##STR00076##
PPDA-020 ##STR00077## PPDA-021 ##STR00078## PPDA-022 ##STR00079##
PPDA-023 ##STR00080## PPDA-024 ##STR00081## PPDA-025 ##STR00082##
PPDA-026 ##STR00083## PPDA-027 ##STR00084## PPDA-028 ##STR00085##
PPDA-029 ##STR00086## PPDA-030 ##STR00087## PPDA-031 ##STR00088##
PPDA-032 ##STR00089##
[0404] Combinations
[0405] It is appreciated that certain features of the invention,
which are, for clarity, described in the context of separate
embodiments, may also be provided in combination in a single
embodiment. Conversely, various features of the invention, which
are, for brevity, described in the context of a single embodiment,
may also be provided separately or in any suitable sub-combination.
All combinations of the embodiments pertaining to the chemical
groups represented by the variables (e.g., --R.sup.2, --R.sup.3,
--R.sup.5X, --R.sup.5Y, --R.sup.6, --R.sup.7, -L.sup.5X-,
-L.sup.5XA-, --R.sup.L5X, -Q, n, -J, m, --R.sup.Q, --R.sup.J,
-L.sup.J-, --R.sup.QA, --R.sup.5YA, --R.sup.7X, --R.sup.7A,
--R.sup.7B, --R.sup.7C, --R.sup.7D, --R.sup.7E, - L.sup.7-,
--W.sup.1, --W.sup.2, --W.sup.3, --R.sup.W1, --R.sup.W11,
--R.sup.W2, --R.sup.W22, --R.sup.W3, --R.sup.W33, --R.sup.3A,
--R.sup.3B, --R.sup.2A, --R.sup.2AA, --R.sup.6A, --R.sup.6AA, etc.)
are specifically embraced by the present invention and are
disclosed herein just as if each and every combination was
individually and explicitly disclosed, to the extent that such
combinations embrace compounds that are stable compounds (i.e.,
compounds that can be isolated, characterised, and tested for
biological activity). In this context, the skilled person will
readily appreciate that certain combinations of groups (e.g.,
substituents) may give rise to compounds which which may not be
readily synthesized and/or are chemically unstable. In addition,
all sub-combinations of the chemical groups listed in the
embodiments describing such variables are also specifically
embraced by the present invention and are disclosed herein just as
if each and every such sub-combination of chemical groups was
individually and explicitly disclosed herein.
[0406] Substantially Purified Forms
[0407] One aspect of the present invention pertains to PPDA
compounds, as described herein, in substantially purified form
and/or in a form substantially free from contaminants.
[0408] In one embodiment, the substantially purified form is at
least 50% by weight, e.g., at least 60% by weight, e.g., at least
70% by weight, e.g., at least 80% by weight, e.g., at least 90% by
weight, e.g., at least 95% by weight, e.g., at least 97% by weight,
e.g., at least 98% by weight, e.g., at least 99% by weight.
[0409] Unless otherwise specified, the substantially purified form
refers to the compound in any stereoisomeric or enantiomeric form.
For example, in one embodiment, the substantially purified form
refers to a mixture of stereoisomers, i.e., purified with respect
to other compounds. In one embodiment, the substantially purified
form refers to one stereoisomer, e.g., optically pure stereoisomer.
In one embodiment, the substantially purified form refers to a
mixture of enantiomers. In one embodiment, the substantially
purified form refers to a equimolar mixture of enantiomers (i.e., a
racemic mixture, a racemate). In one embodiment, the substantially
purified form refers to one enantiomer, e.g., optically pure
enantiomer.
[0410] In one embodiment, the contaminants represent no more than
50% by weight, e.g., no more than 40% by weight, e.g., no more than
30% by weight, e.g., no more than 20% by weight, e.g., no more than
10% by weight, e.g., no more than 5% by weight, e.g., no more than
3% by weight, e.g., no more than 2% by weight, e.g., no more than
1% by weight.
[0411] Unless specified, the contaminants refer to other compounds,
that is, other than stereoisomers or enantiomers. In one
embodiment, the contaminants refer to other compounds and other
stereoisomers. In one embodiment, the contaminants refer to other
compounds and the other enantiomer.
[0412] In one embodiment, the substantially purified form is at
least 60% optically pure (i.e., 60% of the compound, on a molar
basis, is the desired stereoisomer or enantiomer, and 40% is the
undesired stereoisomer or enantiomer), e.g., at least 70% optically
pure, e.g., at least 80% optically pure, e.g., at least 90%
optically pure, e.g., at least 95% optically pure, e.g., at least
97% optically pure, e.g., at least 98% optically pure, e.g., at
least 99% optically pure.
[0413] Isomers
[0414] Certain compounds may exist in one or more particular
geometric, optical, enantiomeric, diastereoisomeric, epimeric,
atropic, stereoisomeric, tautomeric, conformational, or anomeric
forms, including but not limited to, cis- and trans-forms; E- and
Z-forms; c-, t-, and r-forms; endo- and exo-forms; R--, S--, and
meso-forms; D- and L-forms; d- and I-forms; (+) and (-) forms;
keto-, enol-, and enolate-forms; syn- and anti-forms; synclinal-
and anticlinal-forms; .alpha.- and .beta.-forms; axial and
equatorial forms; boat-, chair-, twist-, envelope-, and
halfchair-forms; and combinations thereof, hereinafter collectively
referred to as "isomers" (or "isomeric forms").
[0415] A reference to a class of structures may well include
structurally isomeric forms falling within that class (e.g.,
C.sub.1-7alkyl includes n-propyl and iso-propyl; butyl includes n-,
iso-, sec-, and tert-butyl; methoxyphenyl includes ortho-, meta-,
and para-methoxyphenyl). However, reference to a specific group or
substitution pattern is not intended to include other structural
(or constitutional isomers) which differ with respect to the
connections between atoms rather than by positions in space. For
example, a reference to a methoxy group, --OCH.sub.3, is not to be
construed as a reference to its structural isomer, a hydroxymethyl
group, --CH.sub.2OH. Similarly, a reference specifically to
ortho-chlorophenyl is not to be construed as a reference to its
structural isomer, meta-chlorophenyl.
[0416] The above exclusion does not pertain to tautomeric forms,
for example, keto-, enol-, and enolate-forms, as in, for example,
the following tautomeric pairs: keto/enol (illustrated below),
imine/enamine, amide/imino alcohol, amidine/amidine, nitroso/oxime,
thioketone/enethiol, N-nitroso/hydroxyazo, and nitro/aci-nitro. A
reference herein to one tautomer is intended to encompass both
tautomers.
##STR00090##
[0417] For example, 1H-pyridin-2-one-5-yl and
2-hydroxyl-pyridin-5-yl (shown below) are tautomers of one another.
A reference herein to one is intended to encompass both.
##STR00091##
[0418] Note that specifically included in the term "isomer" are
compounds with one or more isotopic substitutions. For example, H
may be in any isotopic form, including .sup.1H, .sup.2H (D), and
.sup.3H (T); C may be in any isotopic form, including .sup.12C,
.sup.13C, and .sup.14C; O may be in any isotopic form, including
.sup.16O and .sup.18O; and the like.
[0419] Unless otherwise specified, a reference to a particular
compound includes all such isomeric forms, including mixtures
(e.g., racemic mixtures) thereof. Methods for the preparation
(e.g., asymmetric synthesis) and separation (e.g., fractional
crystallisation and chromatographic means) of such isomeric forms
are either known in the art or are readily obtained by adapting the
methods taught herein, or known methods, in a known manner.
[0420] Salts
[0421] It may be convenient or desirable to prepare, purify, and/or
handle a corresponding salt of the compound, for example, a
pharmaceutically-acceptable salt. Examples of pharmaceutically
acceptable salts are discussed in Berge et al., 1977,
"Pharmaceutically Acceptable Salts," J. Pharm. Sci., Vol. 66, pp.
1-19.
[0422] For example, if the compound is anionic, or has a functional
group, which may be anionic (e.g., --COOH may be --COO.sup.-), then
a salt may be formed with a suitable cation. Examples of suitable
inorganic cations include, but are not limited to, alkali metal
ions such as Na.sup.+ and K.sup.+, alkaline earth cations such as
Ca.sup.2+ and Mg.sup.2+, and other cations such as Al.sup.3+ as
well as the ammonium ion (i.e., NH.sub.4.sup.+). Examples of
suitable organic cations include, but are not limited to
substituted ammonium ions (e.g., NH.sub.3R.sup.+,
NH.sub.2R.sub.2.sup.+, NHR.sub.3.sup.+, NR.sub.4.sup.+), for
example, where each R is independently linear or branched saturated
C.sub.1-18alkyl, C.sub.3-8cycloalkyl,
C.sub.3-8cycloalkyl-C.sub.1-6alkyl, and phenyl-C.sub.1-6alkyl,
wherein the phenyl group is optionally substituted. Examples of
some suitable substituted ammonium ions are those derived from:
ethylamine, diethylamine, dicyclohexylamine, triethylamine,
butylamine, ethylenediamine, ethanolamine, diethanolamine,
piperazine, benzylamine, phenylbenzylamine, choline, meglumine, and
tromethamine, as well as amino acids, such as lysine and arginine.
An example of a common quaternary ammonium ion is
N(CH.sub.3).sub.4.sup.+.
[0423] If the compound is cationic, or has a functional group,
which upon protonation may become cationic (e.g., --NH.sub.2 may
become --NH.sub.3.sup.+), then a salt may be formed with a suitable
anion.
[0424] For example, if a parent structure contains a cationic group
(e.g., --NMe.sub.2.sup.+), or has a functional group, which upon
protonation may become cationic (e.g., --NH.sub.2 may become
--NH.sub.3.sup.+), then a salt may be formed with a suitable anion.
In the case of a quaternary ammonium compound a counter-anion is
generally always present in order to balance the positive charge.
If, in addition to a cationic group (e.g., --NMe.sub.2.sup.+,
--NH.sub.3.sup.+), the compound also contains a group capable of
forming an anion (e.g., --COOH), then an inner salt (also referred
to as a zwitterion) may be formed.
[0425] Examples of suitable inorganic anions include, but are not
limited to, those derived from the following inorganic acids:
hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfurous, nitric,
nitrous, phosphoric, and phosphorous.
[0426] Examples of suitable organic anions include, but are not
limited to, those derived from the following organic acids:
2-acetyloxybenzoic, acetic, trifluoroacetic, ascorbic, aspartic,
benzoic, camphorsulfonic, cinnamic, citric, edetic,
1,2-ethanedisulfonic, ethanesulfonic, fumaric, glucoheptonic,
gluconic, glutamic, glycolic, hydroxymaleic, hydroxynaphthalene
carboxylic, isethionic, lactic, lactobionic, lauric, maleic, malic,
methanesulfonic, mucic, oleic, oxalic, palmitic, pamoic,
pantothenic, phenylacetic, phenylsulfonic, propionic, pyruvic,
salicylic, stearic, succinic, sulfanilic, tartaric,
toluenesulfonic, and valeric. Examples of suitable polymeric
organic anions include, but are not limited to, those derived from
the following polymeric acids: tannic acid, carboxymethyl
cellulose.
[0427] Examples of suitable counter-ions which are especially
suitable for quaternary ammonium compounds (e.g., those with a
--NMe.sub.2.sup.+ group) include 1-adamantanesulfonate,
benzenesulfonate, bisulfate, bromide, chloride, iodide,
methanesulfonate, methylsulfate, 1,5-napthalene-bis-sulfonate,
4-nitrobenzenesulfonate, formate, tartrate, tosylate,
trifluoroacetate, trifluoromethylsulfonate, sulphate. Again, if the
compound also contains a group capable of forming an anion (e.g.,
--COOH), then an inner salt may be formed.
[0428] Unless otherwise specified, a reference to a particular
compound also includes salt forms thereof.
[0429] Solvates and Hydrates
[0430] It may be convenient or desirable to prepare, purify, and/or
handle a corresponding solvate of the compound. The term "solvate"
is used herein in the conventional sense to refer to a complex of
solute (e.g., compound, salt of compound) and solvent. If the
solvent is water, the solvate may be conveniently referred to as a
hydrate, for example, a mono-hydrate, a di-hydrate, a tri-hydrate,
etc.
[0431] Unless otherwise specified, a reference to a particular
compound also includes solvate and hydrate forms thereof.
[0432] Chemically Protected Forms
[0433] It may be convenient or desirable to prepare, purify, and/or
handle the compound in a chemically protected form. The term
"chemically protected form" is used herein in the conventional
chemical sense and pertains to a compound in which one or more
reactive functional groups are protected from undesirable chemical
reactions under specified conditions (e.g., pH, temperature,
radiation, solvent, and the like). In practice, well-known chemical
methods are employed to reversibly render unreactive a functional
group, which otherwise would be reactive, under specified
conditions. In a chemically protected form, one or more reactive
functional groups are in the form of a protected or protecting
group (alternatively as a masked or masking group or a blocked or
blocking group). By protecting a reactive functional group,
reactions involving other unprotected reactive functional groups
can be performed, without affecting the protected group; the
protecting group may be removed or the masking group transformed,
usually in a subsequent step, without substantially affecting the
remainder of the molecule. See, for example, Protective Groups in
Organic Synthesis (T. Green and P. Wuts; 4th Edition; John Wiley
and Sons, 2006).
[0434] A wide variety of such "protecting," "blocking," or
"masking" methods are widely used and well known in organic
synthesis. For example, a compound which has two nonequivalent
reactive functional groups, both of which would be reactive under
specified conditions, may be derivatized to render one of the
functional groups "protected," and therefore unreactive, under the
specified conditions; so protected, the compound may be used as a
reactant which has effectively only one reactive functional group.
After the desired reaction (involving the other functional group)
is complete, the protected group may be "deprotected" to return it
to its original functionality.
[0435] For example, a hydroxy group may be protected as an ether
(--OR) or an ester (--OC(.dbd.O)R), for example, as: a t-butyl
ether; a benzyl, benzhydryl (diphenylmethyl), or trityl
(triphenylmethyl) ether; a trimethylsilyl or t-butyldimethylsilyl
ether; or an acetyl ester (--OC(.dbd.O)CH.sub.3, --OAc).
[0436] For example, an aldehyde or ketone group may be protected as
an acetal (R--CH(OR).sub.2) or ketal (R.sub.2C(OR).sub.2),
respectively, in which the carbonyl group (>O.dbd.O) is
converted to a 1,1-diether (>C(OR).sub.2), by reaction with, for
example, a primary alcohol in the presence of an acid. The aldehyde
or ketone group is readily regenerated, for example, by hydrolysis
using water in the presence of acid.
[0437] For example, an amine group may be protected, for example,
as an amide (--NRCO--R) or a urethane (--NRCO--OR), for example,
as: an acetamide (--NHCO--CH.sub.3); a benzyloxy amide
(--NHCO--OCH.sub.2C.sub.6H.sub.5, --NH-Cbz); as a t-butoxy amide
(--NHCO--OC(CH.sub.3).sub.3, --NH-Boc); a 2-biphenyl-2-propoxy
amide (--NHCO--OC(CH.sub.3).sub.2C.sub.6H.sub.4C.sub.6H.sub.5,
--NH-Bpoc), as a 9-fluorenylmethoxy amide (--NH-Fmoc), as a
6-nitroveratryloxy amide (--NH-Nvoc), as a 2-trimethylsilylethyloxy
amide (--NH-Teoc), as a 2,2,2-trichloroethyloxy amide (--NH-Troc),
as an allyloxy amide (--NH-Alloc), as a 2(-phenylsulfonyl)ethyloxy
amide (--NH-Psec); or, in suitable cases (e.g., cyclic amines), as
a nitroxide radical (>N--O ).
[0438] For example, a carboxylic acid group may be protected as an
ester for example, as: an C.sub.1-7alkyl ester (e.g., a methyl
ester; a t-butyl ester); a C.sub.1-7haloalkyl ester (e.g., a
2,2,2-trihaloethyl ester); a 2-tri(C.sub.1-7alkyl)silyl-ethyl
ester; or a C.sub.5-20aryl-C.sub.1-7alkyl ester (e.g., a benzyl
ester; a nitrobenzyl ester); or as an amide or hydrazide, for
example, as acetamide or a N,N,N'-trimethylhydrazide.
[0439] For example, a thiol group may be protected as a thioether
(--SR), for example, as: a benzyl thioether; an acetamidomethyl
ether (--S--CH.sub.2NHC(.dbd.O)CH.sub.3).
[0440] Prodrugs
[0441] It may be convenient or desirable to prepare, purify, and/or
handle the compound in the form of a prodrug. The term "prodrug,"
as used herein, pertains to a compound, which yields the desired
active compound in vivo. Typically, the prodrug is inactive, or
less active than the desired active compound, but may provide
advantageous handling, administration, or metabolic properties.
[0442] For example, some prodrugs are esters of the active compound
(e.g., a physiologically acceptable metabolically labile ester).
During metabolism, the ester group (--C(.dbd.O)OR) is cleaved to
yield the active drug. Such esters may be formed by esterification,
for example, of any of the carboxylic acid groups (--C(.dbd.O)OH)
in the parent compound, with, where appropriate, prior protection
of any other reactive groups present in the parent compound,
followed by deprotection if required.
[0443] Also, some prodrugs are activated enzymatically to yield the
active compound, or a compound, which, upon further chemical
reaction, yields the active compound (for example, as in antibody
directed enzyme prodrug therapy (ADEPT), gene directed enzyme
prodrug therapy (GDEPT), lipid directed enzyme prodrug therapy
(LIDEPT), etc.). For example, the prodrug may be a sugar derivative
or other glycoside conjugate, or may be an amino acid ester
derivative.
[0444] Compositions
[0445] One aspect of the present invention pertains to a
composition (e.g., a pharmaceutical composition) comprising a PPDA
compound, as described herein, and a pharmaceutically acceptable
carrier, diluent, or excipient.
[0446] Another aspect of the present invention pertains to a method
of preparing a composition (e.g., a pharmaceutical composition)
comprising mixing a PPDA compound, as described herein, and a
pharmaceutically acceptable carrier, diluent, or excipient.
[0447] Uses
[0448] The PPDA compounds described herein are useful in the
treatment of, for example, proliferative disorders (as
"anti-proliferative agents"), cancer (as "anti-cancer agents"),
viral infections (as "anti-viral agents"), neurodegenerative
diseases (as "anti-neurodegenerative agents"), etc.
[0449] Use in Methods of Inhibiting CDK
[0450] One aspect of the present invention pertains to a method of
inhibiting CDK (e.g., CDK1, CDK2, CDK4, CDK5, CDK6, CDK7, CDK8,
CDK9, CDK10, CDK11, CDK12, CDK13, etc.) function (e.g., in a cell),
in vitro or in vivo, comprising contacting the cell with an
effective amount of a PPDA compound, as described herein.
[0451] One of ordinary skill in the art is readily able to
determine whether or not a candidate compound inhibits CDK (e.g.,
CDK1, CDK2, CDK4, CDK5, CDK6, CDK7, CDK8, CDK9, CDK10, CDK11,
CDK12, CDK13, etc.). For example, suitable assays are described
herein or are known in the art.
[0452] In one embodiment, the method is performed in vitro.
[0453] In one embodiment, the method is performed in vivo.
[0454] In one embodiment, the PPDA compound is provided in the form
of a pharmaceutically acceptable composition.
[0455] Any type of cell may be treated, including adipose, lung,
gastrointestinal (including, e.g., bowel, colon), breast (mammary),
ovarian, prostate, liver (hepatic), kidney (renal), bladder,
pancreas, brain, and skin.
[0456] For example, a sample of cells may be grown in vitro and a
compound brought into contact with said cells, and the effect of
the compound on those cells observed. As an example of "effect,"
the morphological status of the cells (e.g., alive or dead, etc.)
may be determined. Where the compound is found to exert an
influence on the cells, this may be used as a prognostic or
diagnostic marker of the efficacy of the compound in methods of
treating a patient carrying cells of the same cellular type.
[0457] Use in Methods of Inhibiting Cell Proliferation, etc.
[0458] The PPDA compounds described herein, e.g., (a) regulate
(e.g., inhibit) cell proliferation; (b) inhibit cell cycle
progression; (c) promote apoptosis; or (d) a combination of one or
more of these.
[0459] One aspect of the present invention pertains to a method of
regulating (e.g., inhibiting) cell proliferation (e.g.,
proliferation of a cell), inhibiting cell cycle progression,
promoting apoptosis, or a combination of one or more these, in
vitro or in vivo, comprising contacting a cell with an effective
amount of a PPDA compound, as described herein.
[0460] In one embodiment, the method is a method of regulating
(e.g., inhibiting) cell proliferation (e.g., proliferation of a
cell), in vitro or in vivo, comprising contacting a cell with an
effective amount of a PPDA compound, as described herein.
[0461] In one embodiment, the method is performed in vitro.
[0462] In one embodiment, the method is performed in vivo.
[0463] In one embodiment, the PPDA compound is provided in the form
of a pharmaceutically acceptable composition.
[0464] Any type of cell may be treated, including lung,
gastrointestinal (including, e.g., bowel, colon), breast (mammary),
ovarian, prostate, liver (hepatic), kidney (renal), bladder,
pancreas, brain, and skin.
[0465] One of ordinary skill in the art is readily able to
determine whether or not a candidate compound regulates (e.g.,
inhibits) cell proliferation, etc. For example, assays, which may
conveniently be used to assess the activity offered by a particular
compound are described herein.
[0466] For example, a sample of cells (e.g., from a tumour) may be
grown in vitro and a compound brought into contact with said cells,
and the effect of the compound on those cells observed. As an
example of "effect," the morphological status of the cells (e.g.,
alive or dead, etc.) may be determined. Where the compound is found
to exert an influence on the cells, this may be used as a
prognostic or diagnostic marker of the efficacy of the compound in
methods of treating a patient carrying cells of the same cellular
type.
[0467] Use in Methods of Therapy
[0468] Another aspect of the present invention pertains to a PPDA
compound, as described herein, for use in a method of treatment of
the human or animal body by therapy, for example, for use a method
of treatment of a disorder (e.g., a disease) as described
herein.
[0469] Use in the Manufacture of Medicaments
[0470] Another aspect of the present invention pertains to use of a
PPDA compound, as described herein, in the manufacture of a
medicament, for example, for use in a method of treatment, for
example, for use a method of treatment of a disorder (e.g., a
disease) as described herein.
[0471] In one embodiment, the medicament comprises the PPDA
compound.
[0472] Methods of Treatment
[0473] Another aspect of the present invention pertains to a method
of treatment, for example, a method of treatment of a disorder
(e.g., a disease) as described herein, comprising administering to
a subject in need of treatment a therapeutically-effective amount
of a PPDA compound, as described herein, preferably in the form of
a pharmaceutical composition.
[0474] Disorders Treated
[0475] In one embodiment (e.g., for use in methods of therapy, of
use in the manufacture of medicaments, of methods of treatment),
the treatment is treatment of: a disorder (e.g., a disease) that is
associated with CDK; a disorder (e.g., a disease) resulting from an
inappropriate activity of a CDK; a disorder (e.g., a disease) that
is associated with CDK mutation; a disorder (e.g., a disease) that
is associated with CDK overexpression; a disorder (e.g., a disease)
that is associated with upstream pathway activation of CDK; a
disorder (e.g., a disease) that is ameliorated by the inhibition
(e.g., selective inhibition) of CDK.
[0476] In one embodiment (e.g., for use in methods of therapy, of
use in the manufacture of medicaments, of methods of treatment),
the treatment is treatment of: a proliferative disorder; cancer; a
viral infection (e.g., HIV); a neurodegenerative disorder (e.g.,
Alzheimer's disease, Parkinson's disease); ischaemia; a renal
disease; or a cardiovascular disorder (e.g., atherosclerosis).
[0477] Disorders Treated-Disorders Associated with CDK
[0478] In one embodiment (e.g., for use in methods of therapy, of
use in the manufacture of medicaments, of methods of treatment),
the treatment is treatment of a disorder (e.g., a disease) that is
associated with CDK (e.g., CDK1, CDK2, CDK4, CDK5, CDK6, CDK7,
CDK8, CDK9, CDK10, CDK11, CDK12, CDK13, etc.).
[0479] In one embodiment, the treatment is treatment of: a disorder
(e.g., a disease) resulting from an inappropriate activity of a CDK
(e.g., CDK1, CDK2, CDK4, CDK5, CDK6, CDK7, CDK8, CDK9, CDK10,
CDK11, CDK12, CDK13, etc.).
[0480] In one embodiment, the treatment is treatment of: a disorder
(e.g., a disease) that is associated with CDK mutation; CDK
overexpression (e.g., as compared to corresponding normal cells;
e.g., wherein the overexpression is by a factor of 1.5, 2, 3, 5,
10, 20 or 50); or upstream pathway activation of CDK.
[0481] In one embodiment, the treatment is treatment of a disorder
(e.g., a disease) that is ameliorated by the inhibition (e.g.,
selective inhibition) of CDK (e.g., CDK1, CDK2, CDK4, CDK5, CDK6,
CDK7, CDK8, CDK9, CDK10, CDK11, CDK12, CDK13, etc.).
[0482] Disorders Treated-Proliferative Disorders
[0483] In one embodiment (e.g., for use in methods of therapy, of
use in the manufacture of medicaments, of methods of treatment),
the treatment is treatment of a proliferative disorder.
[0484] The term "proliferative disorder," as used herein, pertains
to an unwanted or uncontrolled cellular proliferation of excessive
or abnormal cells which is undesired, such as neoplastic or
hyperplastic growth.
[0485] In one embodiment, the treatment is treatment of: a
proliferative disorder characterised by benign, pre-malignant, or
malignant cellular proliferation.
[0486] In one embodiment, the treatment is treatment of:
hyperplasia; a neoplasm; a tumour (e.g., a histocytoma, a glioma,
an astrocyoma, an osteoma); cancer; psoriasis; a bone disease; a
fibroproliferative disorder (e.g., of connective tissues);
pulmonary fibrosis; atherosclerosis; or smooth muscle cell
proliferation in the blood vessels (e.g., stenosis or restenosis
following angioplasty).
[0487] Disorders Treated-Cancer
[0488] In one embodiment (e.g., of use in methods of therapy, of
use in the manufacture of medicaments, of methods of treatment),
the treatment is treatment of cancer.
[0489] In one embodiment, the treatment is treatment of cancer
metastasis.
[0490] Included among cancers are:
[0491] (1) Carcinomas, including tumours derived from stratified
squamous epithelia (squamous cell carcinomas) and tumours arising
within organs or glands (adenocarcinomas). Examples include breast,
colon, lung, prostate, ovary.
[0492] (2) Sarcomas, including: osteosarcoma and osteogenic sarcoma
(bone); chondrosarcoma (cartilage); leiomyosarcoma (smooth muscle);
rhabdomyosarcoma (skeletal muscle); mesothelial sarcoma and
mesothelioma (membranous lining of body cavities); fibrosarcoma
(fibrous tissue); angiosarcoma and haemangioendothelioma (blood
vessels); liposarcoma (adipose tissue); glioma and astrocytoma
(neurogenic connective tissue found in the brain); myxosarcoma
(primitive embryonic connective tissue); mesenchymous and mixed
mesodermal tumour (mixed connective tissue types).
[0493] (3) Myeloma.
[0494] (4) Haematopoietic tumours, including: myelogenous and
granulocytic leukaemia (malignancy of the myeloid and granulocytic
white blood cell series); lymphatic, lymphocytic, and lymphoblastic
leukaemia (malignancy of the lymphoid and lymphocytic blood cell
series); polycythaemia vera (malignancy of various blood cell
products, but with red cells predominating).
[0495] (5) Lymphomas, including: Hodgkin and Non-Hodgkin
lymphomas.
[0496] (6) Mixed Types, including, e.g., adenosquamous carcinoma;
mixed mesodermal tumour; carcinosarcoma; teratocarcinoma.
[0497] For example, in one embodiment, the treatment is treatment
of breast cancer.
[0498] In one embodiment, the cancer is characterised by, or
further characterised by, cancer stem cells.
[0499] In one embodiment, the cancer is associated with CDK (e.g.,
CDK1, CDK2, CDK4, CDK5, CDK6, CDK7, CDK8, CDK9, CDK10, CDK11,
CDK12, CDK13, etc.), especially CDK7.
[0500] In one embodiment, the cancer is characterised by, or
further characterised by, inappropriate activity of CDK (e.g.,
CDK1, CDK2, CDK4, CDK5, CDK6, CDK7, CDK8, CDK9, CDK10, CDK11,
CDK12, CDK13, etc.), especially CDK7.
[0501] In one embodiment, the cancer is characterised by, or
further characterised by, overexpression of CDK (e.g., CDK1, CDK2,
CDK4, CDK5, CDK6, CDK7, CDK8, CDK9, CDK10, CDK11, CDK12, CDK13,
etc.), especially CDK7.
[0502] The anti-cancer effect may arise through one or more
mechanisms, including but not limited to, the regulation of cell
proliferation, the inhibition of cell cycle progression, the
inhibition of angiogenesis (the formation of new blood vessels),
the inhibition of metastasis (the spread of a tumour from its
origin), the inhibition of cell migration (the spread of cancer
cells to other parts of the body), the inhibition of invasion (the
spread of tumour cells into neighbouring normal structures), the
promotion of apoptosis (programmed cell death), death by necrosis,
or induction of death by autophagy. The compounds described herein
may be used in the treatment of the cancers described herein,
independent of the mechanisms discussed herein.
[0503] Disorders Treated-Viral Infection
[0504] In one embodiment (e.g., for use in methods of therapy, of
use in the manufacture of medicaments, of methods of treatment),
the treatment is treatment of a viral infection.
[0505] In one embodiment, the treatment is treatment of a viral
infection by:
[0506] (Group I:) a dsDNA virus, e.g., an adenovirus, a
herpesvirus, a poxvirus;
[0507] (Group II:) a ssDNA virus, e.g., a parvovirus;
[0508] (Group III:) a dsRNA virus, e.g., a reovirus;
[0509] (Group IV:) a (+)ssRNA virus, e.g., a picornavirus, a
togavirus;
[0510] (Group V:) a (-)ssRNA virus, e.g., an orthomyxovirus, a
rhabdovirus;
[0511] (Group VI:) a ssRNA-RT virus, e.g., a retrovirus; or
[0512] (Group VII:) a dsDNA-RT virus, e.g., a hepadnavirus.
[0513] As used above: ds: double strand; ss: +strand; (+)ssRNA:
+strand RNA; (-)ssRNA: -strand RNA; ssRNA-RT: (+strand)RNA with DNA
intermediate in life-cycle.
[0514] In one embodiment, the treatment is treatment of: human
immunodeficiency virus (HIV); hepatitis B virus (HBV); hepatitis C
virus (HCV); human papilloma virus (HPV); cytomegalovirus (CMV); or
Epstein-Barr virus (EBV); human herpesvirus 8 (HHV) associated with
Kaposi sarcoma; Coxsackievirus B3; Borna virus; influenza
virus.
[0515] Treatment
[0516] The term "treatment," as used herein in the context of
treating a disorder, pertains generally to treatment of a human or
an animal (e.g., in veterinary applications), in which some desired
therapeutic effect is achieved, for example, the inhibition of the
progress of the disorder, and includes a reduction in the rate of
progress, a halt in the rate of progress, alleviation of symptoms
of the disorder, amelioration of the disorder, and cure of the
disorder. Treatment as a prophylactic measure (i.e., prophylaxis)
is also included. For example, use with patients who have not yet
developed the disorder, but who are at risk of developing the
disorder, is encompassed by the term "treatment."
[0517] For example, treatment includes the prophylaxis of cancer,
reducing the incidence of cancer, alleviating the symptoms of
cancer, etc.
[0518] The term "therapeutically-effective amount," as used herein,
pertains to that amount of a compound, or a material, composition
or dosage form comprising a compound, which is effective for
producing some desired therapeutic effect, commensurate with a
reasonable benefit/risk ratio, when administered in accordance with
a desired treatment regimen.
[0519] Combination Therapies
[0520] The term "treatment" includes combination treatments and
therapies, in which two or more treatments or therapies are
combined, for example, sequentially or simultaneously. For example,
the compounds described herein may also be used in combination
therapies, e.g., in conjunction with other agents. Examples of
treatments and therapies include chemotherapy (the administration
of active agents, including, e.g., drugs, antibodies (e.g., as in
immunotherapy), prodrugs (e.g., as in photodynamic therapy, GDEPT,
ADEPT, etc.); surgery; radiation therapy; photodynamic therapy;
gene therapy; and controlled diets.
[0521] One aspect of the present invention pertains to a compound
as described herein, in combination with one or more (e.g., 1, 2,
3, 4, etc.) additional therapeutic agents, as described below.
[0522] The particular combination would be at the discretion of the
physician who would select dosages using his common general
knowledge and dosing regimens known to a skilled practitioner.
[0523] The agents (i.e., the compound described herein, plus one or
more other agents) may be administered simultaneously or
sequentially, and may be administered in individually varying dose
schedules and via different routes. For example, when administered
sequentially, the agents can be administered at closely spaced
intervals (e.g., over a period of 5-10 minutes) or at longer
intervals (e.g., 1, 2, 3, 4 or more hours apart, or even longer
periods apart where required), the precise dosage regimen being
commensurate with the properties of the therapeutic agent(s).
[0524] The agents (i.e., the compound described here, plus one or
more other agents) may be formulated together in a single dosage
form, or alternatively, the individual agents may be formulated
separately and presented together in the form of a kit, optionally
with instructions for their use.
[0525] Examples of additional agents/therapies that may be
co-administered/combined with treatment with the PPDA compounds
described herein include the following: [0526] an aromatase
inhibitor, for example, exemestane (also known as Aromasin),
letrozole (also known as Femara), anastrozole (also known as
Arimidex), etc.; [0527] an anti-estrogen, for example, faslodex
(also known as Fulvestrant and ICI1182780), tamoxifen (also known
as Nolvadex), hydroxytamoxifen, etc.; [0528] a Her2 blocker, for
example, herceptin, pertuzumab, lapatinib, etc.; a cytotoxic
chemotherapeutic agent, for example, a taxane (e.g., paclitaxel
also known as Taxol; docetaxel also known as Taxotere),
cyclophosphamide, an antimetabolite (e.g., carboplatin,
capecitabine, gemcitabine, doxorubicin, epirubicin, 5-fluorouracil,
etc.), etc.
[0529] Thus, in one embodiment, the treatment further comprises
treatment (e.g., simultaneous or sequential treatment) with a
further active agent which is, e.g., an aromatase inhibitor, an
anti-estrogen, a Her2 blocker, a cytotoxic chemotherapeutic agent,
etc.
[0530] Combination Therapy with an Aromatase Inhibitor and/or an
Anti-Estrogen
[0531] Estrogen receptor a (ERa) is expressed in 70% of breast
tumours and is recognised as the major driver of breast cancer
development and progression in these cases.
[0532] Consequently, ER.alpha. is the predominant target for
adjuvant therapies in ER.alpha.-positive breast cancer. Inhibition
of its activity with anti-estrogens and by inhibition of estrogen
biosynthesis (e.g., using aromatase inhibitors), reduces relapse
and improves patient survival (see, e.g., Osborne, 1998; Cuzick et
al., 2010). Tamoxifen (Nolvadex) is an anti-estrogen that acts by
competing with estrogen for binding to the estrogen receptor, to
inhibit ER.alpha. activity. Importantly, many patients with
ER.alpha.-positive breast cancer relapse on these hormonal
therapies, resistant tumours mostly remaining ER.alpha.-positive
(see, e.g., Ali et al., 2002; Johnston et al., 2003; Ali et al,
2011; Osborne et al., 2011).
[0533] Tamoxifen is an exemplifier of the class of anti-estrogen
known as selective estrogen receptor modulators (SERMs), which are
anti-estrogenic in the breast, but often have estrogen-like
activities in other tissues, such as the cardiovascular system, and
bone. Tamoxifen has been used widely as first line adjuvant agent
for the treatment of ER.alpha.-positive breast cancer in pre- and
post-menopausal women. Fulvestrant (Faslodex) is an anti-estrogen
that competes with estrogen for binding to ER.alpha. to prevent its
activation, but also promotes down-regulation of the ER.alpha.
protein. As such, fulvestrant is an exempifier of the class of
anti-estrogens known as selective estrogen recepto downregulators
(SERD). Fulvestrant is primarily used in the treatment of
ER.alpha.-positive breast cancer patients who experience recurrence
following treatment with first-line adjuvant agents such as
tamoxifen.
[0534] Aromatase is a cytochrome P450 enzyme that catalyses the
limiting step in conversion of androgens to estrogens. Clinically,
anastrozole (Arimidex) and letrozole (Femara) are competitive
inhibitors of the aromatase complex, whilst exemestane (Aromasin)
is an irreversible inhibitor of aromatase. Aromatse inhibitors act
by inhibiting estrogan biosynthesis and thereby levels of
circulating estrogens and consequently by limiting estrogen
availability they prevent ER.alpha. activation.
[0535] Estrogen binding to ER.alpha. protein occurs in the ligand
(hormone) binding domain (LBD), which is C-terminal to the DNA
binding domain (DBD), to promote ER.alpha. dimerisation, nuclear
localisation and binding to DNA in regulatory regions of target
genes, to regulate the expression of said target genes.
Phosphorylation of ER.alpha. provides a key mechanism for
regulating ER.alpha. activity, including DNA binding and
transcription activation. In particular, ER.alpha. phosphorylation
at serine-118 in a region N-terminal to the DBD that is important
for transcription activation by ER.alpha. (known as transcription
activation function 1 (AF-1), is one of the earlies events in
ER.alpha. activation. Serine-118 phophorylation is mediated by
estrogen stimulated recruitment of the transcription factor
complex, TFIIH, which includes CDK7. Estrogen-stimulated TFIIH
recruitment to the estrogen-bound LBD allows CDK7-mediated
phosphorylation of serine-118, to promote ER.alpha. activity. CDK7
over-expression can promote ER.alpha. activity under conditions of
low estrogen levels, as engendered by aromatse inhibitors, and lead
to activation of the tamoxifen-bound ER.alpha. (see, e.g., Ali et
al., 1993; Chen et al., 2000; Chen et al., 2002).
[0536] These findings provide the basis for the use of a PPDA
compound, as described herein, in combination with an aromatase
inhibitor or an anti-estrogen, for the treatment of breast cancer
patients. Such a combination therapy would be especially useful in
the treatment of breast cancer patients following emergence of
resistance to the aromatase inhibitor or anti-estrogen. Such a
combination therapy would also permit the use of reduced amounts
and/or concentrations of the PPDA compound, the anti-estrogen,
and/or the aromatase inhibitor, in order to reduce toxicity.
[0537] Studies demonstrating the synergistic effects of the
combination of a particular PPDA compound (PPDA-001, also referred
to herein as ICEC0942) with an anti-estrogen (4-hydroxytamoxifen or
Faslodex) in the estrogen-responsive ER.alpha.-positive MCF-7
breast cancer cell line are described below. The agents acts
co-operatively to inhibit the growth of breast cancer cells.
[0538] MCF-7 cells purchased from ATCC (USA) were routinely
passaged in DMEM, supplemented with 10% fetal calf serum (FCS) and
kept in a 37.degree. C. incubator with 5% CO.sub.2. The growth
assay for both cell lines was performed in the appropriate media
using using the exact same protocol as described here. For the
growth assay, 5000 cells were seeded into each well of 96-well
plates in DMEM containing 10% FCS. MCF-7 cells were grown over a
12-day period in the presence of PPDA-001/ICEC0942 (100 nmol/L) and
4-hydroxytamoxifen (1 nmol/L) or Faslodex (1 nmol/L). Vehicle
refers to the addition of an equal volume of the solvent (ethanol)
in which 4-hydroxytamoxifen (100 nmol/L) and Faslodex (100 nmol/L)
were prepared. 4-hydroxytamoxifen and Faslodex were prepared at a
concentration of 100 .mu.mol/L in ethanol, and so were diluted in
culture medium at 1 .mu.L per 1 mL of medium (1:1000 dilution), to
give a final concentration of 100 nmol/L. At each time point,
plates were removed for an SRB assay. For this assay, cells were
fixed by the addition of 100 .mu.L/well of ice-cold 40%
trichloroacetic acid (TCA). The plates were left for 1 hour at
4.degree. C., washed in water, and then 100 .mu.L of 0.4% (w/v)
sulforhodamine (SRB; Sigma-Aldrich, UK) prepared in 1% acetic acid
was added. Plates were washed in 1% acetic acid to remove excess
SRB reagent, air dried, and bound dye was solubilized by the
addition of 100 .mu.L of 10 mM tris base. The plates were read at
492 nm using a plate reader. Relative growth was plotted relative
to the OD492 reading for cells at treatment time t=0 (n=5).
[0539] The results for 4-hydroxytamoxifen are illustrated in FIG.
4, which is a graph of relative growth (mean growth.+-.standard
errors of the mean) as function of time for treatment with: (a)
vehicle; (b) PPDA-001/ICEC0942; (c) 4-hydroxytamoxifen; and (d)
PPDA-001/ICEC0942 with 4-hydroxytamoxifen. As shown, co-treatment
greatly enhances the growth inhibition observed for each drug
alone.
[0540] The results for Faslodex are illustrated in FIG. 5, which is
a graph of relative growth (mean growth.+-.standard errors of the
mean) as function of time for treatment with: (a) vehicle; (b)
PPDA-001/ICEC0942; (c) Faslodex; and (d) PPDA-001/ICEC0942 with
Faslodex. As shown, co-treatment greatly enhances the growth
inhibition observed for each drug alone.
[0541] MCF-7 cells cultured in DMEM supplemented with 10% FCS were
treated with 1 .mu.mol/L PPDA-001/ICEC0942 for the time period
shown. As PPDA-001/ICEC0942 was solubilized in DMSO, an equal
volume of DMSO was added as a control. Cell lysates were prepared
by the addition of RIPA buffer (Sigma-Aldrich, cat no: R0278).
Immunoblotting was carried out using standard methods (see, e.g.,
Harlow & Lane, 1988, Antibodies: A Laboratory Manual, Cold
Spring Harbor Laboratory Press, USA), with antibodies for the
protein or phosphorylation mark indicated. .beta.-actin was used as
control for protein loading. The various primary antibodies and
their dilutions are shown in the following table. The second
antibody was HRP-coupled Goat anti Mouse or Rabbit IgG.
TABLE-US-00003 Company/ 1.degree. Antibody 2.degree. Antibody
Antibody Catalog Number Dilution Dilution ER.alpha. (6F11) Vector
1:1000 1:2500 Laboratories (VP-E613) ER.alpha. (phospho-Ser118)
Santa Cruz 1:500 1:2500 (SC-101675) RNA Poll II Abcam 1:500 1:2500
(phospho-Ser2) (Ab5095) RNA Poll II Abcam 1:500 1:2500
(phospho-Ser5) (Ab5131) RNA Poll II Abcam 1:500 1:2500 (Ab5408)
CDK2 (phospho-T160) Abanova 1:500 1:2500 (PAB0438) CDK2 Cell
signaling 1:500 1:2500 (2546) CDK1 (phospho-T161) Cell Signaling
1:500 1:2500 (9114) CDK1 Cell Signaling 1:500 1:2500 (9112)
.beta.-Actin Abcam 1:500 1:2500 (Ab2380)
[0542] FIG. 6 shows immunoblot gels for the bands for
phospho-ER.sup.S118, ER, and .beta.-actin, as a function of time,
for cell lysates from MCF-7 cells treated with 1 .mu.mol/L
PPDA-001/ICEC0942. As shown in the figure, PPDA-001/ICEC0942
inhibits phosphorylation of ER at serine-118
(phospho-ER.sup.S118).
[0543] FIG. 7 shows immunoblot gels for cell lysates from MCF-7
cells treated for 24 hours with 10 .mu.mol/L PPDA-001/ICEC0942.
[0544] Thus, in one embodiment, the treatment further comprises
treatment (e.g., simultaneous or sequential treatment) with a
further active agent which is an aromatase inhibitor, for example,
exemestane (also known as Aromasin), letrozole (also known as
Femara), or anastrozole (also known as Arimidex). In one
embodiment, the disorder is breast cancer (e.g., breast cancer
which is resistant to said aromatase inhibitor).
[0545] Also, in one embodiment, the treatment further comprises
treatment (e.g., simultaneous or sequential treatment) with a
further active agent which is an anti-estrogen, for example,
faslodex (also known as Fulvestrant and ICI182780), tamoxifen (also
known as Nolvadex), or hydroxytamoxifen. In one embodiment, the
disorder is breast cancer (e.g., breast cancer which is resistant
to said anti-estrogen).
[0546] Other Uses
[0547] The PPDA compounds described herein may also be used as cell
culture additives to inhibit CDK (e.g., CDK1, CDK2, CDK4, CDK5,
CDK6, CDK7, CDK8, CDK9, CDK10, CDK11, CDK12, CDK13, etc.).
[0548] The PPDA compounds described herein may also be used as part
of an in vitro assay, for example, in order to determine whether a
candidate host is likely to benefit from treatment with the
compound in question.
[0549] The PPDA compounds described herein may also be used as a
standard, for example, in an assay, in order to identify other
active compounds, other CDK (e.g., CDK1, CDK2, CDK4, CDK5, CDK6,
CDK7, CDK8, CDK9, CDK10, CDK11, CDK12, CDK13, etc.) inhibitors,
etc.
[0550] Kits
[0551] One aspect of the invention pertains to a kit comprising (a)
a PPDA compound as described herein, or a composition comprising a
PPDA compound as described herein, e.g., preferably provided in a
suitable container and/or with suitable packaging; and (b)
instructions for use, e.g., written instructions on how to
administer the compound or composition.
[0552] The written instructions may also include a list of
indications for which the active ingredient is a suitable
treatment.
[0553] Routes of Administration
[0554] The PPDA compound or pharmaceutical composition comprising
the PPDA compound may be administered to a subject by any
convenient route of administration, whether
systemically/peripherally or topically (i.e., at the site of
desired action).
[0555] Examples of routes of administration include oral (e.g., by
ingestion); buccal; sublingual; transdermal (including, e.g., by a
patch, plaster, etc.); transmucosal (including, e.g., by a patch,
plaster, etc.); intranasal (e.g., by nasal spray); ocular (e.g., by
eyedrops); pulmonary (e.g., by inhalation or insufflation therapy
using, e.g., via an aerosol, e.g., through the mouth or nose);
rectal (e.g., by suppository or enema); vaginal (e.g., by pessary);
parenteral, for example, by injection, including subcutaneous,
intradermal, intramuscular, intravenous, intraarterial,
intracardiac, intrathecal, intraspinal, intracapsular, subcapsular,
intraorbital, intraperitoneal, intratracheal, subcuticular,
intraarticular, subarachnoid, and intrasternal; by implant of a
depot or reservoir, for example, subcutaneously or
intramuscularly.
[0556] The Subiect/Patient
[0557] The subject/patient may be a chordate, a vertebrate, a
mammal, a placental mammal, a marsupial (e.g., kangaroo, wombat), a
rodent (e.g., a guinea pig, a hamster, a rat, a mouse), murine
(e.g., a mouse), a lagomorph (e.g., a rabbit), avian (e.g., a
bird), canine (e.g., a dog), feline (e.g., a cat), equine (e.g., a
horse), porcine (e.g., a pig), ovine (e.g., a sheep), bovine (e.g.,
a cow), a primate, simian (e.g., a monkey or ape), a monkey (e.g.,
marmoset, baboon), an ape (e.g., gorilla, chimpanzee, orangutang,
gibbon), or a human.
[0558] Furthermore, the subject/patient may be any of its forms of
development, for example, a foetus.
[0559] In one preferred embodiment, the subject/patient is a
human.
[0560] Formulations
[0561] While it is possible for a PPDA compound to be administered
alone, it is preferable to present it as a pharmaceutical
formulation (e.g., composition, preparation, medicament) comprising
at least one PPDA compound, as described herein, together with one
or more other pharmaceutically acceptable ingredients well known to
those skilled in the art, including pharmaceutically acceptable
carriers, diluents, excipients, adjuvants, fillers, buffers,
preservatives, anti-oxidants, lubricants, stabilisers,
solubilisers, surfactants (e.g., wetting agents), masking agents,
colouring agents, flavouring agents, and sweetening agents. The
formulation may further comprise other active agents, for example,
other therapeutic or prophylactic agents.
[0562] Thus, the present invention further provides pharmaceutical
compositions, as defined above, and methods of making a
pharmaceutical composition comprising mixing at least one PPDA
compound, as described herein, together with one or more other
pharmaceutically acceptable ingredients well known to those skilled
in the art, e.g., carriers, diluents, excipients, etc. If
formulated as discrete units (e.g., tablets, etc.), each unit
contains a predetermined amount (dosage) of the compound.
[0563] The term "pharmaceutically acceptable," as used herein,
pertains to compounds, ingredients, materials, compositions, dosage
forms, etc., which are, within the scope of sound medical judgment,
suitable for use in contact with the tissues of the subject in
question (e.g., human) without excessive toxicity, irritation,
allergic response, or other problem or complication, commensurate
with a reasonable benefit/risk ratio. Each carrier, diluent,
excipient, etc. must also be "acceptable" in the sense of being
compatible with the other ingredients of the formulation.
[0564] Suitable carriers, diluents, excipients, etc. can be found
in standard pharmaceutical texts, for example, Remington's
Pharmaceutical Sciences, 18th edition, Mack Publishing Company,
Easton, Pa., 1990; and Handbook of Pharmaceutical Excipients, 5th
edition, 2005.
[0565] The formulations may be prepared by any methods well known
in the art of pharmacy. Such methods include the step of bringing
into association the compound with a carrier, which constitutes one
or more accessory ingredients. In general, the formulations are
prepared by uniformly and intimately bringing into association the
compound with carriers (e.g., liquid carriers, finely divided solid
carrier, etc.), and then shaping the product, if necessary.
[0566] The formulation may be prepared to provide for rapid or slow
release; immediate, delayed, timed, or sustained release; or a
combination thereof.
[0567] Formulations may suitably be in the form of liquids,
solutions (e.g., aqueous, non-aqueous), suspensions (e.g., aqueous,
non-aqueous), emulsions (e.g., oil-in-water, water-in-oil),
elixirs, syrups, electuaries, mouthwashes, drops, tablets
(including, e.g., coated tablets), granules, powders, losenges,
pastilles, capsules (including, e.g., hard and soft gelatin
capsules), cachets, pills, ampoules, boluses, suppositories,
pessaries, tinctures, gels, pastes, ointments, creams, lotions,
oils, foams, sprays, mists, or aerosols.
[0568] Formulations may suitably be provided as a patch, adhesive
plaster, bandage, dressing, or the like which is impregnated with
one or more compounds and optionally one or more other
pharmaceutically acceptable ingredients, including, for example,
penetration, permeation, and absorption enhancers. Formulations may
also suitably be provided in the form of a depot or reservoir.
[0569] The compound may be dissolved in, suspended in, or mixed
with one or more other pharmaceutically acceptable ingredients. The
compound may be presented in a liposome or other microparticulate
which is designed to target the compound, for example, to blood
components or one or more organs.
[0570] Formulations suitable for oral administration (e.g., by
ingestion) include liquids, solutions (e.g., aqueous, non-aqueous),
suspensions (e.g., aqueous, non-aqueous), emulsions (e.g.,
oil-in-water, water-in-oil), elixirs, syrups, electuaries, tablets,
granules, powders, capsules, cachets, pills, ampoules, boluses.
[0571] Formulations suitable for buccal administration include
mouthwashes, losenges, pastilles, as well as patches, adhesive
plasters, depots, and reservoirs. Losenges typically comprise the
compound in a flavored basis, usually sucrose and acacia or
tragacanth. Pastilles typically comprise the compound in an inert
matrix, such as gelatin and glycerin, or sucrose and acacia.
Mouthwashes typically comprise the compound in a suitable liquid
carrier.
[0572] Formulations suitable for sublingual administration include
tablets, losenges, pastilles, capsules, and pills.
[0573] Formulations suitable for oral transmucosal administration
include liquids, solutions (e.g., aqueous, non-aqueous),
suspensions (e.g., aqueous, non-aqueous), emulsions (e.g.,
oil-in-water, water-in-oil), mouthwashes, losenges, pastilles, as
well as patches, adhesive plasters, depots, and reservoirs.
[0574] Formulations suitable for non-oral transmucosal
administration include liquids, solutions (e.g., aqueous,
non-aqueous), suspensions (e.g., aqueous, non-aqueous), emulsions
(e.g., oil-in-water, water-in-oil), suppositories, pessaries, gels,
pastes, ointments, creams, lotions, oils, as well as patches,
adhesive plasters, depots, and reservoirs.
[0575] Formulations suitable for transdermal administration include
gels, pastes, ointments, creams, lotions, and oils, as well as
patches, adhesive plasters, bandages, dressings, depots, and
reservoirs.
[0576] Tablets may be made by conventional means, e.g., compression
or moulding, optionally with one or more accessory ingredients.
Compressed tablets may be prepared by compressing in a suitable
machine the compound in a free-flowing form such as a powder or
granules, optionally mixed with one or more binders (e.g.,
povidone, gelatin, acacia, sorbitol, tragacanth,
hydroxypropylmethyl cellulose); fillers or diluents (e.g., lactose,
microcrystalline cellulose, calcium hydrogen phosphate); lubricants
(e.g., magnesium stearate, talc, silica); disintegrants (e.g.,
sodium starch glycolate, cross-linked povidone, cross-linked sodium
carboxymethyl cellulose); surface-active or dispersing or wetting
agents (e.g., sodium lauryl sulfate); preservatives (e.g., methyl
p-hydroxybenzoate, propyl p-hydroxybenzoate, sorbic acid);
flavours, flavour enhancing agents, and sweeteners. Tablets may be
made by moulding in a suitable machine a mixture of the powdered
compound moistened with an inert liquid diluent. The tablets may
optionally be coated or scored and may be formulated so as to
provide slow or controlled release of the compound therein using,
for example, hydroxypropylmethyl cellulose in varying proportions
to provide the desired release profile. Tablets may optionally be
provided with a coating, for example, to affect release, for
example an enteric coating, to provide release in parts of the gut
other than the stomach.
[0577] Ointments are typically prepared from the compound and a
paraffinic or a water-miscible ointment base.
[0578] Creams are typically prepared from the compound and an
oil-in-water cream base. If desired, the aqueous phase of the cream
base may include, for example, at least about 30% w/w of a
polyhydric alcohol, i.e., an alcohol having two or more hydroxyl
groups such as propylene glycol, butane-1,3-diol, mannitol,
sorbitol, glycerol and polyethylene glycol and mixtures thereof.
The topical formulations may desirably include a compound which
enhances absorption or penetration of the compound through the skin
or other affected areas. Examples of such dermal penetration
enhancers include dimethylsulfoxide and related analogues.
[0579] Emulsions are typically prepared from the compound and an
oily phase, which may optionally comprise merely an emulsifier
(otherwise known as an emulgent), or it may comprise a mixture of
at least one emulsifier with a fat or an oil or with both a fat and
an oil. Preferably, a hydrophilic emulsifier is included together
with a lipophilic emulsifier which acts as a stabiliser. It is also
preferred to include both an oil and a fat. Together, the
emulsifier(s) with or without stabiliser(s) make up the so-called
emulsifying wax, and the wax together with the oil and/or fat make
up the so-called emulsifying ointment base which forms the oily
dispersed phase of the cream formulations.
[0580] Suitable emulgents and emulsion stabilisers include Tween
60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl
monostearate and sodium lauryl sulfate. The choice of suitable oils
or fats for the formulation is based on achieving the desired
cosmetic properties, since the solubility of the compound in most
oils likely to be used in pharmaceutical emulsion formulations may
be very low. Thus the cream should preferably be a non-greasy,
non-staining and washable product with suitable consistency to
avoid leakage from tubes or other containers. Straight or branched
chain, mono- or dibasic alkyl esters such as di-isoadipate,
isocetyl stearate, propylene glycol diester of coconut fatty acids,
isopropyl myristate, decyl oleate, isopropyl palmitate, butyl
stearate, 2-ethylhexyl palmitate or a blend of branched chain
esters known as Crodamol CAP may be used, the last three being
preferred esters. These may be used alone or in combination
depending on the properties required. Alternatively, high melting
point lipids such as white soft paraffin and/or liquid paraffin or
other mineral oils can be used.
[0581] Formulations suitable for intranasal administration, where
the carrier is a liquid, include, for example, nasal spray, nasal
drops, or by aerosol administration by nebuliser, include aqueous
or oily solutions of the compound.
[0582] Formulations suitable for intranasal administration, where
the carrier is a solid, include, for example, those presented as a
coarse powder having a particle size, for example, in the range of
about 20 to about 500 microns which is administered in the manner
in which snuff is taken, i.e., by rapid inhalation through the
nasal passage from a container of the powder held close up to the
nose.
[0583] Formulations suitable for pulmonary administration (e.g., by
inhalation or insufflation therapy) include those presented as an
aerosol spray from a pressurised pack, with the use of a suitable
propellant, such as dichlorodifluoromethane,
trichlorofluoromethane, dichoro-tetrafluoroethane, carbon dioxide,
or other suitable gases.
[0584] Formulations suitable for ocular administration include eye
drops wherein the compound is dissolved or suspended in a suitable
carrier, especially an aqueous solvent for the compound.
[0585] Formulations suitable for rectal administration may be
presented as a suppository with a suitable base comprising, for
example, natural or hardened oils, waxes, fats, semi-liquid or
liquid polyols, for example, cocoa butter or a salicylate; or as a
solution or suspension for treatment by enema.
[0586] Formulations suitable for vaginal administration may be
presented as pessaries, tampons, creams, gels, pastes, foams or
spray formulations containing in addition to the compound, such
carriers as are known in the art to be appropriate.
[0587] Formulations suitable for parenteral administration (e.g.,
by injection) include aqueous or non-aqueous, isotonic,
pyrogen-free, sterile liquids (e.g., solutions, suspensions), in
which the compound is dissolved, suspended, or otherwise provided
(e.g., in a liposome or other microparticulate). Such liquids may
additionally contain other pharmaceutically acceptable ingredients,
such as anti-oxidants, buffers, preservatives, stabilisers,
bacteriostats, suspending agents, thickening agents, and solutes,
which render the formulation isotonic with the blood (or other
relevant bodily fluid) of the intended recipient. Examples of
excipients include, for example, water, alcohols, polyols,
glycerol, vegetable oils, and the like. Examples of suitable
isotonic carriers for use in such formulations include Sodium
Chloride Injection, Ringer's Solution, or Lactated Ringer's
Injection. Typically, the concentration of the compound in the
liquid is from about 1 ng/mL to about 10 .mu.g/mL, for example from
about 10 ng/mL to about 1 .mu.g/mL. The formulations may be
presented in unit-dose or multi-dose sealed containers, for
example, ampoules and vials, and may be stored in a freeze-dried
(lyophilised) condition requiring only the addition of the sterile
liquid carrier, for example water for injections, immediately prior
to use. Extemporaneous injection solutions and suspensions may be
prepared from sterile powders, granules, and tablets.
[0588] Dosage
[0589] It will be appreciated by one of skill in the art that
appropriate dosages of the PPDA compounds, and compositions
comprising the PPDA compounds, can vary from patient to patient.
Determining the optimal dosage will generally involve the balancing
of the level of therapeutic benefit against any risk or deleterious
side effects. The selected dosage level will depend on a variety of
factors including the activity of the particular PPDA compound, the
route of administration, the time of administration, the rate of
excretion of the PPDA compound, the duration of the treatment,
other drugs, compounds, and/or materials used in combination, the
severity of the disorder, and the species, sex, age, weight,
condition, general health, and prior medical history of the
patient. The amount of PPDA compound and route of administration
will ultimately be at the discretion of the physician,
veterinarian, or clinician, although generally the dosage will be
selected to achieve local concentrations at the site of action
which achieve the desired effect without causing substantial
harmful or deleterious side-effects.
[0590] Administration can be effected in one dose, continuously or
intermittently (e.g., in divided doses at appropriate intervals)
throughout the course of treatment. Methods of determining the most
effective means and dosage of administration are well known to
those of skill in the art and will vary with the formulation used
for therapy, the purpose of the therapy, the target cell(s) being
treated, and the subject being treated. Single or multiple
administrations can be carried out with the dose level and pattern
being selected by the treating physician, veterinarian, or
clinician.
[0591] In general, a suitable dose of the PPDA compound is in the
range of about 10 .mu.g to about 250 mg (more typically about 100
.mu.g to about 25 mg) per kilogram body weight of the subject per
day. Where the compound is a salt, an ester, an amide, a prodrug,
or the like, the amount administered is calculated on the basis of
the parent compound and so the actual weight to be used is
increased proportionately.
[0592] Chemical Syntheses
[0593] Melting points were determined using a Reichert-Thermovar
hot-stage apparatus and are uncorrected. IR spectra were recorded
as thin films with the absorption bands reported in wave number
(cm.sup.-1).
[0594] .sup.1H NMR spectra were recorded at 400 MHz or at 500 MHz.
Chemical shifts are reported as .delta.-values in ppm relative to
the CDCl.sub.3 peak (.delta.H 7.26), to the CD.sub.3OD peak
(.delta.H 3.31), and to the DMSO-d.sub.6 peak (.delta.H 2.54).
Coupling constants (J) recorded in Hertz (Hz) and quoted to the
nearest 0.5 Hz.
[0595] Optical Rotations were recorded with a path length of 1 dm,
using the 589.3 mn D-line of sodium. Concentrations (c) are quoted
in g/100 mL.
[0596] All reactions were carried out with magnetic stirring and if
air or moisture sensitive, in flame-dried or oven-dried glassware
under nitrogen or argon. Syringes, used to transfer reagents and
solvents, were purged with nitrogen or argon prior to use. Reaction
temperatures other than room temperature were recorded as the bath
temperature unless otherwise stated.
[0597] All solvents and reagents were used as commercially
supplied, unless otherwise stated. Et.sub.2O, THF, PhMe and
CH.sub.2Cl.sub.2 were redistilled from Na-Ph.sub.2CO,
Na-Ph.sub.2CO, Na and CaH.sub.2 respectively.
[0598] Thin layer chromatography was performed on pre-coated
aluminum backed silica gel F254 glass plates. The chromatogram was
visualized under UV light and/or by staining using aqueous
potassium permanganate or aqueous acidic vanillin followed by
heating with a heat gun.
[0599] Flash column chromatography was performed using silica gel,
particle size 40-63 .mu.m (eluents are given in parenthesis).
[0600] General Synthesis
[0601] The general synthetic route towards the target compounds is
illustrated in the following scheme. The three main side chains
(R.sup.1, R.sup.2 and R.sup.3) can be greatly varied.
##STR00092##
1-Synthesis of the Aromatic Dichloro-Heterocyclic Compound 9
[0602] The synthesis of the dichloride 9 was carried out in a
manner similar to published methods. See, e.g., Jogalekar et al.,
2008.
##STR00093##
2-Synthesis of the Aromatic Cores
##STR00094##
[0603] 2.1-General Procedure A-Substitution of Chloride 9
[0604] A solution containing 3-iso-propyl-5,7-dichloropyrazolo
[1,5-a]pyrimidine 9 (2.17 mmol) and the amine (4.56 mmol) in EtOH
(20 mL) was heated at reflux for 3 h. The reaction mixture was
cooled to room temperature and concentrated in vacuo. The remaining
residue was purified by flash column chromatography (MeOH:EtOAc) to
yield the desired product in analytically pure form.
2.2-Synthesis of the Aromatic Cores
Synthesis 1
N-Benzyl-5-chloro-3-iso-propylpyrazolo[1,5-a]pyrimidin-7-amine
(10)
##STR00095##
[0606] Following general procedure A, 9 (500 mg, 2.17 mmol) and
benzylamine (0.52 mL, 4.78 mmol) were allowed to react in EtOH (20
mL). The title compound was obtained as a white solid (630 mg, 97%)
after flash column chromatography (hexane:EtOAc 6:1).
[0607] M.p.=74.degree. C. (CHCl.sub.3); .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.82 (m, 1H), 7.32 (m, 5H), 7.01 (m, 1H), 5.90
(m, 1H), 4.53 (m, 2H), 3.27 (sept, J=6.9 Hz, 1H), 1.32 (d, J=6.9
Hz, 6H); .sup.13C (CDCl.sub.3, 300 MHz) .delta. 150.1, 146.8,
144.1, 141.5, 135.7, 129.0, 128.1, 127.1, 116.9, 84.6, 46.0, 23.4,
23.3; HRMS (Cl) calc. for C.sub.16H.sub.17ClN.sub.4 (M+H.sup.+):
301.1220, found: 301.1230; Anal. calc. for
C.sub.16H.sub.17ClN.sub.4: C 63.89, H 5.70, N 18.63, found: C
63.95, H 5.78, N 18.59.
Synthesis 2
5-Chloro-3-iso-propyl-N-(pyridin-4-ylmethyl)
pyrazolo[1,5-a]pyrimidin-7-amine (11)
##STR00096##
[0609] Following general procedure A, 9 (460 mg, 2.0 mmol) and
4-picolylamine (0.407 mL, 4.0 mmol) were allowed to react in EtOH
(4 mL). The title compound was obtained as a pale yellow solid (601
mg, 99%) after flash column chromatography (hexane:EtOAc gradient
7:3 to 1:1).
[0610] .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 150.4, 150.0,
146.9, 145.3, 144.2, 141.8, 121.5, 117.2, 84.8, 44.8, 23.4; HRMS
(ESI) calc. for C.sub.15H.sub.16CIN.sub.5 (M+H.sup.+): 302.1185,
found: 302.1172.
Synthesis 3
tert-Butyl 4-((5-chloro-3-iso-propylpyrazolo
[1,5-a]pyrimidin-7-yl)carbamoyl)piperidine-1-carboxylate (12)
##STR00097##
[0612] tert-Butyl-4-carbamoylpiperidine-1-carboxylate (1 g, 4.37
mmol) was dissolved in dry DMF (4 mL) and treated with NaH (60%
wt., 175 mg, 4.37 mmol) at room temperature for 1 h. The mixture
was treated with 9 (1.0 g, 4.37 mmol). The title compound was
obtained as a white solid (1.1 g, 60%) after flash column
chromatography (hexane:EtOAc 8:2).
[0613] IR (neat): v.sub.max=3141, 1705, 1541; .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 9.31 (s, 1H), 7.80 (s, 1H), 7.57 (s, 1H),
4.28-4.07 (m, 2H), 3.26-3.11 (m, 1H), 2.79 (br. s., 2H), 2.70-2.59
(m, 1H), 1.92 (d, J=11.2 Hz, 2H), 1.76-1.64 (m, 2H), 1.40 (s, 9 H),
1.26 (d, J=6.8 Hz, 6H); .sup.13C NMR (101 MHz, CDCl.sub.3)
.delta.=173.1, 170.9, 154.4, 150.1, 143.5, 141.4, 139.5, 118.0,
94.5, 79.7, 60.1, 43.9, 42.6, 28.2, 28.0, 23.3, 23.1.
Synthesis 4
5-Chloro-3-iso-propyl-N-(4-(4-methylpiperazin-1-yl)phenyl)pyrazolo[1,5-a]p-
yrimidin-7-amine (13)
##STR00098##
[0615] NaH (60% wt., 125 mg, 3.14 mmol) was added with stirring to
a solution of 4-(4-methylpiperazin-1-yl)aniline (see, e.g.,
Sengupta et al., 2012) (60 g, 3.14 mmol) in THF (15 mL). After 10
min, a solution of 9 (0.72 g, 3.14 mmol) in DMF (2 mL) was added
and the mixture was heated to 50.degree. C. for 3 h. The title
compound was obtained as a white solid (80 mg, 66%) after flash
column chromatography (CH.sub.2Cl.sub.2/MeOH 9:1).
[0616] R.sub.f=0.36 (9:1 CH.sub.2Cl.sub.2/MeOH); IR (neat):
.nu..sub.max=3324, 1609, 1577 cm.sup.-1; .sup.1H NMR (CDCl.sub.3,
400 MHz) .delta. 7.97 (s, 1H), 7.90 (s, 1H), 7.26-7.22 (m, 2H),
7.02-6.98 (m, 2H), 6.07 (s, 1H), 3.33 (sept, J=6.7 Hz, 1H), 3.28
(dd, J=6.3, 3.9 Hz, 4H), 2.62 (dd, J=6.3, 3.9 Hz, 4H), 2.39 (s,
3H), 1.35 (d, J=6.9 Hz, 6H); HRMS (ESI) Calcd. for
C.sub.20H.sub.25N.sub.6Cl [M+H].sup.+, 385.1907, found
385.1909.
2.3-General Procedure B-Boc Protection
[0617] Boc.sub.2O (0.73 mmol) was added with stirring to a solution
of the amino-heterocycle (0.52 mmol) and 4-dimethylaminopyridine
(DMAP) (0.31 mmol) in dry THF (2 mL) at 0.degree. C. After 16 h at
room temperature, the reaction mixture was diluted with EtOAc,
washed with water twice and saturated aqueous NaHCO.sub.3, dried
over MgSO.sub.4, filtered and concentrated under reduced pressure.
Purification by flash column chromatography (hexane:Et.sub.2O)
afforded the desired product in analytically pure form.
2.4-Synthesis of the Boc-Protected Aromatic Cores
Synthesis 5
tent-Butyl benzyl(5-chloro-3-iso-propyl
pyrazolo[1,5-a]pyrimidin-7-yl)carbamate (14)
##STR00099##
[0619] Following general procedure B, 10 (300 mg, 1 mmol),
Boc.sub.2O (284 mg, 1.3 mmol) and DMAP (24 mg, 0.2 mmol) were
allowed to react in THF (6 mL). The title compound was obtained as
a pale yellow solid (385 mg, 96%) after flash column chromatography
(hexane: EtOAc 20:1).
[0620] M.p. =93-94.degree. C. (EtOAc); IR (neat):
.nu..sub.max=1727, 1612, 1518, 1454, 1154, 699 cm.sup.-1; .sup.13C
NMR (300 MHz, CDCl.sub.3) .delta. 152.6, 147.9, 144.9, 144.0,
142.5, 136.7, 128.5, 127.7, 127.6, 118.2, 106.1, 82.9, 51.3, 27.8,
23.5, 23.3; HRMS (Cl) calc. for C.sub.21H.sub.25ClN.sub.4O.sub.2
(M+H.sup.+): 401.1744, found: 401.1747; Anal. calc. for
C.sub.21H.sub.25ClN.sub.4O.sub.2: C 62.91, H 6.29, N 13.98, found:
C 62.87, H 6.19, N 13.94.
Synthesis 6
tent-Butyl (5-chloro-3-iso-propylpyrazolo
[1,5-a]pyrimidin-7-yl)(pyridin-4-ylmethyl) carbamate (15)
##STR00100##
[0622] Following general procedure B, 11 (560 mg, 1.85 mmol),
Boc.sub.2O (565 mg, 2.59 mmol) and DMAP (67 mg, 0.55 mmol) were
allowed to react in THF (10 mL). The title compound was obtained as
a pale yellow solid (688 mg, 92%) after flash column chromatography
(hexane: EtOAc 4:1).
[0623] IR (neat): .nu..sub.max=1724, 1610, 1560, 1516, 1367, 1305,
1150, 1103, 877, 730 cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.51 (d, J=5.0 Hz, 2H), 7.97 (s, 1H), 7.20 (d, J=5.0 Hz,
2H), 6.57 (s, 1H), 4.98 (s, 2H), 3.25 (sept, J=6.9 Hz, 1H), 1.32
(s, 9H), 1.30 (d, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3)
.delta. 152.2, 149.8, 147.7, 145.8, 144.9, 143.8, 142.5, 121.9,
118.2, 105.3, 83.2, 50.6, 27.6, 23.4, 23.1; HRMS (ESI) calc. for
C.sub.20H.sub.24ClN.sub.5O.sub.2 (M+H.sup.+): 402.1698, found:
402.1697.
Synthesis 7
tert-Butyl
5-chloro-3-iso-propylpyrazolo[1,5-a]pyrimidin-7-yl(4-(4-methylp-
iperazin-1-yl)phenyl)carbamate (16)
##STR00101##
[0625] Following general procedure B, 13 (0.67 g, 1.74 mmol),
Boc.sub.2O (0.76 g, 3.48 mmol) and DMAP (21 mg, 0.174 mmol) were
allowed to react in 1,4-dioxane (8 mL). The title compound was
obtained as a pale yellow solid (806 mg, 96%) after flash column
chromatography (CH.sub.2Cl.sub.2:MeOH 20:1).
[0626] R.sub.f=0.50 (9:1 CH.sub.2Cl.sub.2/MeOH); IR (neat):
.nu..sub.max=1735, 1608, 1511, 1147 cm.sup.-1; .sup.13C NMR
(CDCl.sub.3, 100 MHz) .delta. 152.1, 150.7, 148.1, 145.3, 145.1,
142.8, 131.4, 127.6, 118.2, 116.1, 105.5, 83.1, 55.0, 48.7, 46.2,
27.9, 23.6, 23.5; HRMS (ESI) Calcd. for
C.sub.25H.sub.33N.sub.6O.sub.2Cl [M+H].sup.+, 485.2432, found
485.2421.
3-Synthesis of the Pyrrolidine and Piperidine Intermediates
3.1-Synthesis of Pyrrolidine 22
##STR00102##
[0628] Synthesis 8
(3S,4S)-1-Benzyl-3,4-dihydroxybyrrolidine-2,5-dione (18)
##STR00103##
[0630] Benzylamine (3.7 mL, 33 mmol) and D-(+)-tartaric acid 17 (5
g, 33 mmol) in xylene (25 mL) were heated at reflux (160.degree.
C.) under vigorously stirring for 6 h using a Dean-Stark apparatus.
The reaction mixture was cooled to room temperature, filtered and
the residue was washed with acetone. Recrystallization from
absolute ethanol (35 mL) gave the title compound (6 g, 81%) as a
white solid. See, e.g., Nagel et al., 1984.
[0631] [.alpha.].sup.20.sub.D=-139 (c 1.0, MeOH); m.p.=198.degree.
C.; .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.24-7.35 (m, 5H),
6.31 (d, J=5.4 Hz, 2H), 4.58 (d, J=14.8 Hz, 1H), 4.52 (d, J=14.8
Hz, 1H), 4.38 (d, J=5.4 Hz, 2H); .sup.13C NMR (100 MHz,
DMSO-d.sub.6) .delta. 174.7, 136.1, 128.7, 127.6, 74.6, 41.3.
Synthesis 9
(3R,4R)-tert-Butyl 3,4-dihydroxypyrrolidine-1-carboxylate (19)
##STR00104##
[0633] LiAlH.sub.4 (2 M in THF, 18.5 mL, 37 mmol) was slowly added
with stirring to a solution of 18 (3 g, 13.6 mmol) in THF (90 mL)
at 0.degree. C. The reaction mixture was heated at reflux for 16 h,
cooled to 0.degree. C. and slowly quenched with H.sub.2O (1.48 mL),
aqueous NaOH (15% wt. in H.sub.2O, 1.48 mL) and H.sub.2O (4.5 mL).
The reaction mixture was filtered through Celite, washed with hot
THF (50 mL) and the filtrate was concentrated in vacuo to give
crude (3R,4R)-1-benzyl-3,4-dihydroxypyrrolidine as a pale yellow
solid, which was used for the next step without further
purification.
[0634] The crude benzylpyrrolidine in MeOH (35 ml) was allowed to
react with Boc2O (3.26 g, 15 mmol) and Pd/C (10% wt., 300 mg) and
stirred under an atmosphere of hydrogen for 16 h. The mixture was
filtered through Celite, concentrated under reduced pressure and
the residue was recrystallized from hot EtOAc (25 mL) yielding 19
(2.0 g, 72%) as pale yellow crystals. See, e.g., Nagel et al.,
1984.
[0635] R.sub.f=0.22 (CH.sub.2Cl.sub.2:MeOH 19:1); .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 3.94 (brt, J=3.3 Hz, 2H), 3.67 (dd,
J=12.0, 4.9 Hz, 1H), 3.63 (dd, J=12.0, 5.0 Hz, 1H), 3.42 (dd,
J=12.0, 2.3 Hz, 2H), 3.37 (dd, J=12.1, 2.3 Hz, 2H), 1.44 (s, 9H);
.sup.13C NMR (400 MHz, CDCl.sub.3) .delta. 153.9, 80.5, 64.2, 63.4,
48.8, 48.5, 28.4; MS (ESI): m/z 204 (M+H.sup.+).
Synthesis 10
(3R,4R)-tert-Butyl
3-hydroxy-4-(methoxy-methoxy)-pyrrolidine-1-carboxylate (20)
##STR00105##
[0637] NaH (60% wt., 5.87 mmol, 235 mg) and MOMCl (445 .mu.L, 5.87
mmol) were added with stirring to a solution of 19 (993 mg, 4.89
mmol) in dry THF (25 mL) at 0.degree. C. The solution was allowed
to reach room temperature and stirred for 5 h. Saturated aqueous
NH.sub.4Cl (10 mL) was added and the aqueous layer was extracted
with EtOAc (3.times.10 mL). The combined organic layers were washed
with brine, dried over MgSO.sub.4, filtered and concentrated under
reduced pressure. The residue was purified by flash column
chromatography (CH.sub.2Cl.sub.2:MeOH 19:1) to give the title
compound (910 mg, 75%) as a pale yellow oil.
[0638] R.sub.f=0.42 (CH.sub.2Cl.sub.2:MeOH 19:1);
[.alpha.].sup.20.sub.D=-45 (c 0.88, CH.sub.2Cl.sub.2); .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta.4.68 (d, J=7.2 Hz, 1H), 4.66 (d, J=7.6
Hz, 1H), 4.18 (dd, J=9.6, 4.0 Hz, 1H), 3.94 (brs, 1H), 3.63 (brs,
2H), 3.38 (s, 3H), 3.26-3.35 (m, 2H), 1.43 (s, 9H); .sup.13C NMR
(100 MHz, CDCl.sub.3) .delta. 154.7, 96.1, 82.3, 79.7, 74.1, 55.8,
51.3, 49.6, 28.6; HRMS (Cl) calc. for C.sub.11H.sub.21NO.sub.5
(M+H.sup.+): 248.1498, found: 248.1494; Anal. calc. for
C.sub.11H.sub.21NO.sub.5: C 53.43, H 8.56, N 5.66, found: C 53.56,
H 8.62, N 5.70.
Synthesis 11
(3S,4R)-tert-Butyl
3-azido-4-(methoxy-methoxy)-pyrrolidine-1-carboxylate (21)
##STR00106##
[0640] Et.sub.3N (1.05 mL, 7.5 mmol) and methanesulfonyl chloride
(440 .mu.L, 5.6 mmol) were added with stirring to a solution of 20
(920 mg, 3.72 mmol) in CH.sub.2Cl.sub.2 (20 mL) at 0.degree. C. The
reaction mixture was stirred at 0.degree. C. for 40 min, quenched
with saturated aqueous NaHCO.sub.3 (10 mL) and extracted with
CH.sub.2Cl.sub.2 (3.times.10 mL). The combined organic layers were
washed with brine, dried over MgSO.sub.4, filtered and concentrated
under reduced pressure. The resulting orange oil was used for the
next step without further purification.
[0641] The crude oil was dissolved in dry DMF (20 mL), sodium azide
(1.2 g, 18.6 mmol) was added and the resulting suspension was
heated at 100.degree. C. for 48 h, cooled to room temperature and
diluted with water (10 mL) and Et.sub.2O (10 mL). The aqueous layer
was extracted with Et.sub.2O (3.times.10 mL) and the combined
organic layers were washed with water and brine, dried over
MgSO.sub.4, filtered and concentrated under reduced pressure. The
residue was purified by flash column chromatography
(hexane:Et.sub.2O 2:3) to yield azide 21 (825 mg, 81%) as a
colorless oil.
[0642] R.sub.f=0.33 (hexane:Et.sub.2O 2:3);
[.alpha.].sup.20.sub.D=-34 (c 1.07, CH.sub.2Cl.sub.2); IR (neat):
.nu..sub.max=2102, 1692, 1399, 1365, 1117, 995 cm.sup.-1; .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 4.73 (d, J=7.0 Hz, 1H), 4.71 (d,
J=7.0 Hz, 1H), 4.25-4.31 (m, 1H), 3.89-3.95 (m, 1H), 3.53-3.60 (m,
2H), 3.37-3.50 (m, 2H), 3.42 (s, 3H), 1.45 (s, 9H); .sup.13C NMR
(100 MHz, CDCl.sub.3) .delta. 154.31, 96.1, 80.0, 76.6, 60.8, 55.9,
49.6, 48.0, 28.5; HRMS (Cl) calc. for
C.sub.11H.sub.20N.sub.4O.sub.5 (M+H.sup.+): 273.1563, found
273.1569; Anal. calc. for C.sub.11H.sub.20N.sub.4O.sub.5: C 48.52,
H 7.40, N 20.58, found: C 48.45, H 7.31, N 20.50.
Synthesis 12
(3S,4R)-tert-Butyl
3-amino-4-(methoxy-methoxy)-pyrrolidine-1-carboxylate (22)
##STR00107##
[0644] Azide 21 (100 mg, 0.37 mmol) in methanol (5 mL) was stirred
and allowed to react with Pd/C (10% wt., 20 mg) under an atmosphere
of hydrogen for 40 min. The mixture was filtered through Celite and
the filtrate was concentrated under reduced pressure. The residue
was purified by flash column chromatography
(CH.sub.2Cl.sub.2:MeOH:30% wt. aqueous ammonia 92.75:7:0.25) to
give 22 (89 mg, 98%) as a colorless oil.
[0645] R.sub.f=0.40 (CH.sub.2Cl.sub.2:MeOH:30% wt aqueous ammonia
92.75:7:0.25); [.alpha.].sup.20.sub.D=-21 (c 1.0,
CH.sub.2Cl.sub.2); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
4.62-4.69 (m, 2H), 3.96 (q, J=7.2 Hz, 1H), 3.48-3.55 (m, 2H),
3.40-3.46 (m, 2H), 3.34 (s, 3H), 2.98-3.10 (m, 1H), 1.55 (brs, 2H),
1.39 (s, 9H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 154.7,
96.0, 79.4, 77.6, 55.7, 53.8, 51.3, 50.0, 28.5; HRMS (Cl) calc. for
C.sub.11H.sub.22N.sub.2O.sub.4 (M+H.sup.-): 247.1658, found
247.1653; Anal. calc. for C.sub.11H.sub.22N.sub.2O.sub.4: C 53.64,
H 9.00, N 11.37, found: C 53.58, H 8.96, N 11.31.
3.2-Synthesis of Amine 31
##STR00108##
[0646] Synthesis 13
(1-Benzyl-1,2,3,6-tetrahydropyridin-4-yl)methanol (24)
##STR00109##
[0648] 4-Pyridinemethanol (25.0 g, 229 mmol) was suspended in MeCN
(250 mL) and benzyl chloride (31.5 ml, 275 mmol) was slowly added.
The reaction was heated at reflux for 3 h, cooled to room
temperature and concentrated under reduced pressure. The red
residue was dissolved in methanol (350 mL) and cooled to
-35.degree. C. Sodium borohydride (17.4 g, 485 mmol) was added
portionwise, maintaining the internal temperature below -20.degree.
C. Once the addition was complete the mixture was stirred for 30
min and quenched by the dropwise addition of water (50 mL). The
mixture was concentrated under reduced pressure, diluted with
CH.sub.2Cl.sub.2 (10 mL) and water (10 mL). The aqueous layer was
extracted with CH.sub.2Cl.sub.2 (3.times.15 mL) and the combined
organic layers were washed with brine, dried over MgSO.sub.4 and
concentrated under reduced pressure. The crude product was purified
by flash column chromatography (hexane:EtOAc gradient 2:3 to 0:1)
to yield 24 (32.0 g, 69%) as a white solid. See, e.g., Gijsen et
al., 2008.
[0649] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.30 (5H, m), 5.54
(1H, m), 4.68 (t, J=5.6 Hz, 1H), 3.83 (brd, J=5.5 Hz, 2H), 3.53
(2H, s), 2.87 (2H, m), 2.50 (t, J=5.7 Hz, 2H), 2.02 (2H, m);
.sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 139.1 137.1, 129.1,
128.6, 127.3, 118.9, 64.7, 62.3, 52.5, 49.8, 26.6.
Synthesis 14
1-Benzyl-4-(hydroxymethyl)piperidine-3,4-diol (25)
##STR00110##
[0651] A pre-mixed clear solution of K.sub.3Fe(CN).sub.6 (4.90 g,
15.0 mmol), K.sub.2CO.sub.3 (2.06 g, 15.0 mmol), (DHQD).sub.2PHAL
(123 mg, 0.16 mmol), K.sub.2OsO.sub.2(OH).sub.4 (29.1 mg, 0.079
mmol) and MeSO.sub.2NH.sub.2 (476 mg, 5.00 mmol) in tBuOH and
H.sub.2O (50 mL, 1:1) was cooled to 0.degree. C. and
tetrahydropyridine 24 (1.02 g, 5.00 mmol) was added. After stirring
for 12 h at 0.degree. C., sodium sulfite was added (30 g) and the
reaction mixture was diluted with H.sub.2O (20 mL). After 1 h
vigorous stirring at room temperature, the reaction mixture was
extracted with CH.sub.2Cl.sub.2 (3.times.10 mL). The combined
organic extracts were dried over MgSO.sub.4 and concentrated under
reduced pressure. Purification by flash column chromatography
(CH.sub.2Cl.sub.2:MeOH: concentrated aqueous NH.sub.3 gradient
98:2:0.5 to 90:10:0.5) afforded 25 (485 mg, 41%) as an orange oil.
See, e.g., Kolb et al., 1994.
[0652] R.sub.f=0.20 (CH.sub.2Cl.sub.2:MeOH 19:1); IR (neat):
.nu..sub.max=3342, 1454, 1300, 1102, 1075, 1045, 1007, 963, 746,
698 cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.36-7.26
(m, 5H), 3.83 (dd, J=8.9, 4.1 Hz, 1H), 3.67-3.51 (m, 4H), 3.49 (s,
1H), 3.16 (brs, 2H), 2.71 (dd, J=10.9, 3.5 Hz, 1H), 2.53-2.34 (m,
3H), 1.69-1.57 (m, 2H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta.
137.3, 129.3, 128.3, 127.3, 71.0, 70.4, 68.7, 62.4, 54.9, 48.4,
31.5; HRMS (ESI) calc. for C.sub.13H.sub.19NO.sub.3 (M+H.sup.+):
238.1443, found: 238.1445.
Synthesis 15
4-(Hydroxymethyl)piperidine-3,4-diol (26)
##STR00111##
[0654] Triol 25 (440 mg, 1.85 mmol) was dissolved in MeOH (3 ml)
and treated with Pd/C (15% wt., 22 mg) and stirred under an
atmosphere of hydrogen for 10 h at 50.degree. C. The mixture was
filtered through Celite and the filtrate was concentrated under
reduced pressure giving amine 26 (270 mg, 99%).
[0655] IR (neat): .nu..sub.max=3273, 1646, 1533, 1420, 1272, 1052,
971, 856, 818 cm.sup.-1; .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
3.61 (dd, J=10.1, 5.0 Hz, 1H), 3.59 (d, J=10.9 Hz, 1H), 3.43 (d,
J=10.9 Hz, 1H), 3.35 (s, 2H), 2.88-2.70 (m, 4H), 1.64-1.61 (m, 2H),
1.69-1.57 (m, 2H); .sup.13C NMR (100 MHz, CD.sub.3OD) .delta. 73.1,
69.8, 67.9, 48.3, 41.7, 33.9; HRMS (Cl) calc. for
C.sub.6H.sub.13NO.sub.3 (M+H.sup.+): 148.0974, found: 148.0974.
Synthesis 16
tert-Butyl-3,4-dihydroxy-4-(hydroxymethyl) piperidine-1-carboxylate
(27)
##STR00112##
[0657] Boc.sub.2O (370 mg, 1.70 mmol) and MeOH (0.5 mL) were added
with stirring to a solution of amine 26 (250 mg, 1.70 mmol) in
CH.sub.2Cl.sub.2 (2 mL) and the reaction mixture was stirred for 3
h at room temperature. The solvent was removed under reduced
pressure and purification by flash column chromatography
(CH.sub.2Cl.sub.2:MeOH gradient 19:1 to 9:1) afforded 27 (342 mg,
81%) of carbamate 27 as a white solid.
[0658] R.sub.f=0.30 (CH.sub.2Cl.sub.2:MeOH 19:1); IR (neat):
.nu..sub.max=3335, 1664, 1425, 1366, 1274, 1250, 1156, 1057, 988,
960 cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 3.98-3.95
(m, 1H), 3.83 (brs, 1H), 3.72 (d, J=11.1 Hz, 1H), 3.57 (d, J=11.1
Hz, 1H), 3.11 (brs, 4H), 2.96 (dd, J=12.4, 10.6 Hz, 1H), 1.68-1.65
(m, 1H), 1.52-1.41 (m, 2H), 1.47 (s, 9H); .sup.13C NMR (100 MHz,
CDCl.sub.3) .delta. 154.9, 80.2, 71.1, 70.5, 69.8, 44.8, 39.0,
31.7, 28.4; HRMS (Cl) calc. for C.sub.11H.sub.21NO.sub.5
(M+H.sup.+): 248.1498, found: 248.1503.
Synthesis 17
tert-Butyl
3,4-dihydroxy-4-((4-toluenesulfonyloxy)-methyl)piperidine-1-car-
boxylate (28)
##STR00113##
[0660] pTsCl (246 mg, 1.29 mmol) was added with stirring to a
solution of triol 27 (290 mg, 1.17 mmol) in dry pyridine (2.4 mL).
The reaction mixture was stirred for 14 h at room temperature,
poured onto water and extracted with CH.sub.2Cl.sub.2 (3.times.3
mL). The combined organic extracts were dried over MgSO.sub.4,
filtered and concentrated under reduced pressure. Purification by
flash column chromatography (hexane:EtOAc gradient 7:3 to 1:1) gave
the sulfonate 28 (237 mg, 50%) as a white solid.
[0661] R.sub.1=0.57 (hexane:EtOAc 2:3); m.p.=110.degree. C.; IR
(neat): .nu.=3411, 1686, 1427, 1359, 1255, 1183, 1169, 1070, 1054,
972, 840, 814, 667 cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.80 (d, J=8.3 Hz, 2H), 7.38 (d, J=8.3 Hz, 2H), 4.07 (d,
J=10.1 Hz, 1H), 4.06 (brs, 1H), 3.90 (d, J=10.1 Hz, 1H), 3.70 (dd,
J=10.5, 5.4 Hz, 1H), 3.89 (brs, 1H), 3.02 (brs, 2H), 2.88 (brs,
1H), 2.61 (brs, 1H), 2.49 (s, 3H), 1.71-1.59 (m, 2H), 1.48 (s, 9H);
.sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 154.6, 145.3, 132.3,
130.0, 128.0, 80.1, 72.7, 70.9, 66.7, 44.1, 38.9, 31.7, 28.4, 21.7;
HRMS (ESI) calc. for C.sub.18H.sub.27NO.sub.7S (M+Na.sup.+):
424.1406, found: 424.1399.
Synthesis 18
tert-Butyl 4-(azidomethyl)-3,4-dihydroxy-piperidine-1-carboxylate
(29)
##STR00114##
[0663] NaN3 (109 mg, 1.68 mmol) was added with stirring to a
solution of 4-toluenesulfonate 28 (225 mg, 0.560 mmol) in dry DMF
(5.6 mL). The reaction mixture was stirred for 12 h at 60.degree.
C., cooled to room temperature and H.sub.2O (25 mL) was added. The
reaction mixture was extracted with Et.sub.2O (3.times.10 mL) and
the combined organic extracts were washed with brine, dried over
MgSO.sub.4, filtered and concentrated under reduced pressure.
Purification by flash column chromatography (hexane:Et.sub.2O 7:3
to 1:4) yielded azide 29 (140 mg, 92%) as a colorless oil.
[0664] R.sub.f=0.29 (hexane:EtOAc 3:2); IR (neat):
.nu..sub.max=3386, 2101, 1664, 1426, 1367, 1275, 1246, 1152, 1068,
873, 763 cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
4.02-4.01 (m, 1H), 3.83 (brs, 1H), 3.63-3.59 (m, 1H), 3.49 (d,
J=12.2 Hz, 1H), 3.37 (d, J=12.2 Hz, 1H), 3.09 (brt, J=11.3 Hz, 1H),
2.99-2.51 (brs, 2H), 2.92 (t, J=11.5 Hz, 1H), 1.79 (d, J=14.0 Hz,
1H), 1.63-1.47 (m, 1H), 1.47 (s, 9H); .sup.13C NMR (100 MHz,
CDCl.sub.3) .delta. 154.8, 80.3, 71.9, 68.2, 58.2, 44.8, 39.2,
32.5, 28.4; HRMS (Cl) calc. for Cu.sub.11H.sub.20N.sub.4O.sub.4
(M+NH.sub.4.sup.+): 290.1828, found: 290.1831.
Synthesis 19
tert-Butyl
7a-(azidomethyl)-2,2-dimethyl-tetrahydro-[1,3]dioxolo[4,5-c]pyr-
idine-5(6H)-carboxylate (30)
##STR00115##
[0666] Azide 29 (130 mg, 0.48 mmol) was dissolved in acetone and
dimethoxypropane (1:1; 6 mL) and pTsOH (9 mg, 0.05 mmol) was added.
The reaction mixture was stirred at room temperature for 2 h and
concentrated under reduced pressure. The residue was purified by
flash column chromatography (hexane:EtOAc 1:1) giving azide 30 (130
mg, 87%) as a colorless oil.
[0667] R.sub.f=0.6 (hexane:EtOAc 1:1); .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 4.20-4.08 (m, 2H), 3.62-3.20 (m, 4H), 3.10-3.02
(m, 1H), 1.88 (m, 1H), 1.51-1.44 (m, 16H); HRMS (ESI) calc. for
C.sub.14H.sub.23N.sub.4O.sub.4 (M+Na.sup.+): 311.1719, found:
311.1702.
Synthesis 20
tert-Butyl
7a-(aminomethyl)-2,2-dimethyl-tetrahydro-[1,3]dioxolo[4,5-c]pyr-
idine-5(6H)-carboxylate (31)
##STR00116##
[0669] Azide 30 (80 mg, 0.26 mmol) was dissolved in MeOH (5 mL),
and stirred and allowed to react with Pd/C (10% wt., 60 mg) under a
hydrogen atmosphere for 3 h. The mixture was filtered through
Celite, washed with MeOH and concentrated under reduced pressure to
afford amine 31 (72 mg, 0.25 mmol, 97%) as a colorless oil which
was used in the next step without further purification.
[0670] R.sub.f=0.25 (CH.sub.2Cl.sub.2:MeOH 19:1).
3.3-Synthesis of Piperidine 38
##STR00117##
[0671] Synthesis 21
1-Benzyl-4-(hydroxymethyl)piperidin-3-ol (32)
##STR00118##
[0673] Tetrahydropyridine 24 (2.00 g, 9.85 mmol) was dissolved in
THF (19 ml) and cooled to -30.degree. C. Borane.THF complex (1 M in
THF, 19.0 ml, 19.0 mmol) was added dropwise and the mixture allowed
to warm to room temperature overnight. The solution was cooled to
-10.degree. C. and quenched by addition of water (0.5 mL). Hydrogen
peroxide (30% in water, 1.24 ml) and sodium hydroxide (3 M in
water, 1.37 ml) were simultaneously added dropwise. Sodium
hydroxide (50% in water, 2.5 ml) was added and the mixture heated
at reflux for 4 h. The reaction mixture was cooled to room
temperature and the white precipitate removed by filtration. The
filtrate was concentrated under reduced pressure, taken up in
CH2Cl.sub.2 and water, poured onto saturated aqueous NaHCO.sub.3
(10 mL). The aqueous layer was extracted with CH.sub.2Cl.sub.2
(3.times.10 mL) and the combined organic layers were washed with
brine, dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The
product was purified by trituration from i-Pr.sub.2O to yield 41
(1.14 g, 53%). See, e.g., Gijsen et al., 2008.
[0674] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.32 (m, 5H), 3.72
(m, 3H), 3.02 (dd, J=10.7, 4.4 Hz, 1H), 2.85 (md, J=11.1 Hz, 1H),
2.63 (s, 1H), 2.19 (brs, 1H), 2.00 (td, J=11.6, 2.5 Hz, 1H), 1.89
(t, J=10.2 Hz, 1H), 1.59 (m, 3H), 1.28 (qd, J=12.8, 4.2 Hz, 1H);
.sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 138.0, 129.2, 128.2,
127.1, 73.1, 68.0, 62.7, 60.0, 52.6, 44.5, 26.1.
Synthesis 22
tert-Butyl 4-(azidomethyl)-3-hydroxy-piperidine-1-carboxylate
(33)
##STR00119##
[0676] Piperidine 32 (2.62 g, 11.9 mmol) and Pd/C (10% wt., 263 mg)
were suspended in MeOH (25.0 ml) and Boc.sub.2O (3.89 g, 17.8 mmol)
was added. The atmosphere was replaced by hydrogen and the mixture
heated to 50.degree. C. overnight. The suspension was cooled to
room temperature, filtered first through Celite and subsequently
through a plug of silica to yield the title compound (2.74 g, 99%),
which was used in the next step without further purification.
[0677] Crude tert-butyl-3-hydroxy-4-(hydroxymethyl)
piperidine-1-carboxylate (414 mg, 1.78 mmol) was dissolved in
CH.sub.2Cl.sub.2 (1 ml) and pyridine (1 ml) and cooled to 0.degree.
C. pTsCl (358 mg, 1.87 mmol) was added portionwise followed by DMAP
(1.00 mg, 0.008 mmol). The mixture was stirred for 48 h, diluted
with CH.sub.2Cl.sub.2 (5 mL) and poured onto hydrochloric acid (0.5
M, 5 mL). The aqueous layer was extracted with CH.sub.2Cl.sub.2
(3.times.3 mL) and the combined organic layers were washed with
brine, dried over MgSO.sub.4 and concentrated under reduced
pressure. The product was purified by flash column chromatography
(hexane:EtOAc gradient 1:9 to 1:1) to yield tert-butyl
3-hydroxy-4-(toluene-4-sulfonyloxymethyl)piperidine-1-carboxylate
(493 mg, 72%), which was used directly without further
purification.
[0678]
tert-butyl-3-hydroxy-4-(toluene-4-sulfonyloxy-methyl)piperidine-1-c-
arboxylate (425 mg, 1.10 mmol) was dissolved in DMF (10 ml) and
NaN.sub.3 (86.0 mg 1.32 mmol) was added. The mixture was heated to
60.degree. C. overnight, cooled to room temperature and poured onto
brine. The aqueous layer was extracted with Et.sub.2O (3.times.3
mL) and the combined organic layers were washed with brine, dried
over MgSO.sub.4 and concentrated under reduced pressure. The
product was purified by flash column chromatography (hexane:EtOAc
gradient 9:1 to 3:1) to yield azide 33 (253 mg, 99%).
[0679] .sup.1H NMR (500 MHz, DMSO-d.sub.6) .delta. 4.58 (brs, 1H),
4.02 (ddd, J=12.9, 5.1, 1.8 Hz, 1H), 3.89 (dddd, J=13.2, 4.5, 2.7,
1.8 Hz, 1H), 3.56 (dd, J=12.4, 3.8 Hz, 1H), 3.35 (dd, J=12.2, 6.8
Hz, 1H), 3.19 (td, J=9.7, 4.8 Hz, 1H), 2.68 (ddd, J=13.2, 12.3, 3.0
Hz, 1H), 2.48 (m, 1H), 1.73 (dq, J=13.3, 3.0 Hz, 1H), 1.56 (m, 1H),
1.42 (s, 9H), 1.22 (qd, J=12.0, 4.5 Hz, 1H); .sup.13C NMR (125 MHz,
DMSO-d.sub.6) .delta. 153.3, 78.0, 66.4, 52.5, 49.4, 42.8, 42.3,
27.5, 26.7.
Synthesis 23
(3R,4R)-tent-Butyl
3-((R)-2-acetoxy-2-phenyl-acetoxy)-4-(azidomethyl)piperidine-1-carboxylat-
e (34)
(3S,4S)-tert-Butyl
3-((R)-2-acetoxy-2-phenyl-acetoxy)-4-(azidomethyl)piperidine-1-carboxylat-
e (35)
##STR00120##
[0681] Azide 33 (7.13 g, 29.0 mmol), (R)-O-acetoxy-mandelic acid
(6.75 g, 34.7 mmol) and DMAP (354 mg, 2.90 mmol) were dissolved in
CH.sub.2Cl.sub.2 (100 mL). EDCl (6.66 g, 34.7 mmol) was added and
the reaction mixture was stirred for 16 h. Reaction was quenched by
addition of water (150 mL). The phases were separated and the
aqueous phase was extracted with CH.sub.2Cl.sub.2 (3.times.100 mL).
The combined organic phases were dried over MgSO.sub.4 and
concentrated under reduced pressure. The diastereoisomeric mixture
was separated by preparative HPLC to afford 34 (5.02 g, 41%) and 35
(5.00 g, 41%) as colorless oils.
[0682] 34: R.sub.f=0.41 (hexane:Et20 1:1);
[.alpha.].sup.25.sub.D=-51.6 (c 2.0, CH.sub.2Cl.sub.3); .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.45-7.44 (m, 2H), 7.40-7.38 (m, 3H),
5.82 (brs, 1H), 4.65 (brs, 1H), 4.02 (brs, 2H), 3.48 (d, 1H, J=11.8
Hz), 3.22 (dd, J=12.4, 7.2 Hz, 1H), 2.64 (t, J=12.5 Hz, 1H), 2.44
(t, J=11.3 Hz, 1H), 2.20 (s, 3H), 1.87-1.77 (m, 2H), 1.41 (s, 10H);
.sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 170.7, 168.3, 154.4,
133.3, 129.6, 129.0, 127.6, 80.4, 74.7, 70.2, 52.6, 46.0, 42.8,
41.3, 28.4, 27.8, 20.8; HRMS (ESI) calc. for
C.sub.21H.sub.28N.sub.4O.sub.6 (M+Na.sup.+): 455.1907, found:
455.1896.
[0683] 35: R.sub.f=0.38 (hexane/Et.sub.2O 1:1);
[.alpha.].sup.25.sub.D=-86.1 (c 2.0, CHCl.sub.3); .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.47-7.45 (m, 2H), 7.40-7.39 (m, 3H), 5.88
(s, 1H), 4.53 (bs, 1H), 4.27 (bs, 1H), 4.01 (bs, 1H), 2.88 (d,
J=11.4 Hz, 1H), 2.72 (dd, J=12.1, 6.9 Hz, 1H), 2.64 (t, J=11.9 Hz,
1H), 2.19 (s, 3H), 1.73-1.59 (m, 3H), 1.44 (s, 9H), 1.31 (dq,
J=12.4, 4.3 Hz, 1H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta.
170.3, 168.0, 154.4, 133.8, 129.6, 129.0, 127.7, 80.3, 74.4, 70.6,
52.1, 46.5, 42.7, 41.0, 28.4, 27.6, 20.8; HRMS (ESI) calc. for
C.sub.21H.sub.28N.sub.4O.sub.6 (M+Na.sup.+): 455.1907, found:
455.1905.
Synthesis 24
(3R,4R)-tert-Butyl
4-(azidomethyl)-3-hydroxypiperidine-1-carboxylate (36)
##STR00121##
[0685] Mandelic ester 34 (5.00 g, 11.6 mmol) and LiOH.H.sub.2O
(1.21 g, 28.9 mmol) were dissolved in THF, MeOH and H.sub.2O
(1:1:1; 60 mL) and stirred for 2 h. The reaction mixture was
diluted with water (100 mL) and extracted with EtOAc (4.times.50
mL). The combined organic phases were dried over MgSO.sub.4 and
concentrated under reduced pressure. The resultant alcohol 36 (2.91
g, 98%) was analytically pure and could be used without further
purification.
[0686] [.alpha.].sup.25.sub.D=+20.1 (c 4.0, MeOH); .sup.1H NMR (500
MHz, DMSO-d.sub.6) .delta. 4.58 (brs, 1H), 4.02 (ddd, J=12.9, 5.1,
1.8 Hz, 1H), 3.89 (dddd, J=13.2, 4.5, 2.7, 1.8 Hz, 1H), 3.56 (dd,
J=12.4, 3.8 Hz, 1H), 3.35 (dd, J=12.2, 6.8 Hz, 1H), 3.19 (td,
J=9.7, 4.8 Hz, 1H), 2.68 (ddd, J=13.2, 12.3, 3.0 Hz, 1H), 2.48 (m,
1H), 1.73 (dq, J=13.3, 3.0 Hz, 1H), 1.56 (m, 1H), 1.42 (s, 9H),
1.22 (dq, J=12.0, 4.5 Hz, 1H); .sup.13C NMR (125 MHz, DMSO-d.sub.6)
.delta. 153.3, 78.0, 66.4, 52.5, 49.4, 42.8, 42.3, 27.5, 26.7.
Synthesis 25
(3R,4R)-tert-Butyl
4-(azidomethyl)-3-(methoxymethoxy)piperidine-1-carboxylate (37)
##STR00122##
[0688] Di-iso-propylethylamine (0.224 mL, 1.29 mmol) and MOMCl (98
.mu.L, 1.29 mmol) were added dropwise with stirring to a solution
of alcohol 36 (165 mg, 0.644 mmol) in CH.sub.2Cl.sub.2 (5 mL) at
0.degree. C. After 14 h at room temperature, the reaction mixture
was poured onto saturated aqueous NH.sub.4Cl (5 mL) and extracted
with Et20 (3.times.2 mL). The combined organic extracts were dried
over MgSO.sub.4, filtered and concentrated under reduced pressure.
Purification by flash column chromatography (hexane:Et.sub.2O
gradient 4:1 to 1:1) to afforded azide 37 (144 mg, 74%) as a
colorless oil.
[0689] R.sub.f=0.69 (hexane:Et.sub.2O 7:3); IR (neat):
.nu..sub.max=3286, 1693, 1421, 1366, 1278, 1153, 1140, 1101, 1037
cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 4.75 (d, J=6.8
Hz, 1H), 4.67 (d, J=6.8 Hz, 1H), 4.39 (brs, 1H), 4.08 (brs, 1H),
3.59 (dd, J=12.2, 3.3 Hz, 1H), 3.43 (s, 3H), 3.43-3.38 (m, 1H),
3.34 (td, J=10.0, 4.9 Hz, 1H), 2.70 (brt, J=12.5 Hz, 1H), 2.55
(brt, J=9.9 Hz, 1H), 1.84-1.79 (m, 1H), 1.76-1.67 (m, 1H), 1.47 (s,
9H), 1.40 (dq, J=12.5, 4.5 Hz, 1H); .sup.13C NMR (100 MHz,
CDCl.sub.3) .delta. 154.5, 96.2, 79.9, 73.9, 55.8, 53.1, 47.7,
43.5, 42.4, 28.4, 28.0; HRMS (Cl) calc. for
C.sub.13H.sub.24N.sub.4O.sub.4 (M+Na.sup.+): 323.1695, found:
323.1693.
Synthesis 26
(3R,4R)-tert-Butyl
4-(aminomethyl)-3-(methoxymethoxy)piperidine-1-carboxylate (38)
##STR00123##
[0691] Pd/C (15% wt., 19.5 mg) was added to a degassed solution of
azide 37 (130 mg, 0.433 mmol) in MeOH (2 mL). After 1.5 h under
hydrogen atmosphere, the reaction mixture was filtered through
Celite and concentrated under reduced pressure. Purification by
flash column chromatography (CH.sub.2Cl.sub.2:MeOH: NH.sub.4OH
gradient 94:5:1 to 87.5:12.5:1) to afforded 38 (98.7 mg, 83%) as a
colorless oil.
[0692] R.sub.f=0.35 (CH.sub.2Cl.sub.2MeOH 9:1); IR (neat):
.nu..sub.max=1689, 1421, 1365, 1244, 1151, 1102, 1030, 917, 882
cm.sup.-1; .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 4.73 (d, J=6.9
Hz, 1H), 4.66 (d, J=6.9 Hz, 1H), 4.29-4.26 (m, 1H), 4.00-3.96 (m,
1H), 3.39 (s, 3H), 3.25 (td, J=9.8, 4.6 Hz, 1H), 2.93 (dd, J=12.8,
4.7 Hz, 1H), 2.78 (brt, J=13.2 Hz, 1H) 2.75-2.56 (m, 1H), 2.58 (dd,
J=12.8, 6.8 H, 1H), 1.84-1.80 (m, 1H), 1.64-1.54 (m, 1H), 1.45 (s,
9H), 1.23 (m, 1H); .sup.13C NMR (100 MHz, CD.sub.3OD) .delta.
156.4, 97.4, 81.3, 77.2, 56.1, 49.5, 45.1, 44.5, 28.7; HRMS (Cl)
calc. for C.sub.13H.sub.26N.sub.2O.sub.4 (M+H.sup.+): 275.1971,
found: 275.1970.
3.4-Synthesis of Piperidine 41
##STR00124##
[0693] Synthesis 27
tent-Butyl 4-hydroxy-4-(nitromethyl)-piperidine-1-carboxylate
(40)
##STR00125##
[0695] tert-Butyl-4-oxopiperidine-1-carboxylate 39 (10.0 g, 50.2
mmol) was added portion-wise with stirring to a solution of
nitromethane (100 mL) and Et.sub.3N (42.8 mL, 301 mmol) and the
mixture was stirred for 4 days at room temperature. The reaction
mixture was diluted with EtOAc (80 mL) and washed with water,
saturated aqueous NH.sub.4Cl and brine, dried over MgSO.sub.4 and
concentrated under reduced pressure to give the title compound
(12.1 g, 93%) as a white solid. See, e.g., Bosmans et al.,
2005.
[0696] R.sub.f=0.10 (hexanes:EtOAc 4:1); m.p.=141.degree. C.; IR
(neat): .nu..sub.max=3383, 1660, 1545 cm.sup.-1; .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 4.42 (s, 2H), 3.93-3.90 (m, 2H),
3.22-3.15 (m, 2H), 2.99 (brs, 1H), 1.68-1.65 (m, 2H), 1.61-1.57 (m,
2H), 1.45 (s, 9H); .sup.13C NMR (100 MHz, CD.sub.3OD) .delta.
154.6, 84.7, 79.9, 69.1, 40.0, 34.2, 28.4; HRMS (ESI) calc. for
C.sub.11H.sub.20N.sub.2O.sub.5 (M+H.sup.+): 260.1367, found:
260.1450.
Synthesis 28
tent-Butyl 4-(aminomethyl)-4-hydroxy-piperidine-1-carboxylate
(41)
##STR00126##
[0698] A stirred solution of piperidine 40 (200 mg, 0.77 mmol) and
imidazole (260 mg, 3.82 mmol) in TESCI (1 mL) and DMF (0.5 mL) was
heated at 70.degree. C. After 20 h, the mixture was cooled to room
temperature and treated with water (100 mL). Following an
extraction with ethyl acetate (3.times.50 mL) the combined organic
phases were washed with brine (1.times.50 mL), dried (MgSO.sub.4)
and concentrated in vacuo to give an oily yellow residue containing
the TES ether intermediate. This material was used in the next step
of the reaction sequence without further purification.
[0699] A stirred solution of the TES ether intermediate, in dry
MeOH (5 mL), was treated with Pd/C (10% wt., 200 mg) and placed
under an atmosphere of hydrogen at 22 bars. After 48 h, the ensuing
mixture was filtered and the filtrate was concentrated in vacuo to
give a yellow oil. Subjection of this material to flash
chromatography (CH.sub.2Cl.sub.2:MeOH gradient 19:1 to 9:1)
afforded the amine 41 (182 mg, 69%) as a colorless oil.
[0700] R.sub.f=0.5 (EtOAc); IR (neat): .nu..sub.max, =3396, 2953,
2913, 2875, 1692, 1421, 1365, 1243, 1155, 1060 cm.sup.-1; .sup.1H
NMR (CDCl.sub.3, 400 MHz) .delta. 3.61-3.57 (m, 2H), 3.28 (ddd,
J=13.2, 8.8, 4.3 Hz, 2H), 2.67 (s, 2H), 1.55-1.51 (m, 2H), 1.43 (s,
9H), 1.17 (s, 2H), 0.94 (t, J=7.9 Hz, 9H), 0.59 (q, J=7.9 Hz, 6H);
.sup.13C NMR (100 MHz, CD.sub.3OD) .delta. 154.8, 79.3, 74.1, 51.5,
40.1, 35.0, 28.4, 7.2, 6.9; HRMS (ESI) calc. for
C.sub.17H.sub.37N.sub.2O.sub.3Si (M+H.sup.+): 345.2573, found:
345.2563.
3.5-Synthesis of Pyrrolidine 46
##STR00127##
[0701] Synthesis 29
tert-Butyl 3,3-bis(hydroxymethyl)-pyrrolidine-1-carboxylate
(43)
##STR00128##
[0703] Pyrrolidine 42 (see, e.g., Xu et al., 2011) (2.62 g, 11.9
mmol) and Pd/C (10% wt., 263 mg) were suspended in MeOH (25.0 mL)
and Boc2O (3.89 g, 17.8 mmol) was added. The atmosphere was
replaced by hydrogen and the mixture heated to 50.degree. C.
overnight. The suspension was cooled to room temperature and
filtered first through Celite and subsequently through a small
silica plug to yield carbamate 43 (2.74 g, 99%).
[0704] R.sub.f=0.43 (CH.sub.2Cl.sub.2:MeOH:saturated aqueous
NH.sub.3 10:1:0.1); IR (neat): .nu..sub.max=3394, 1666, 1610, 1574,
1477, 1415, 1366, 1254, 1149, 1107, 1039, 914, 879, 771, 731, 700,
646 cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 4.06 (brs,
1H), 3.91 (brs, 1H), 3.60 (s, 4H), 3.39-3.32 (m, 2H), 3.20-3.17 (m,
2H), 1.70 (t, J=7.2 Hz, 2H), 1.41 (s, 9H).
Synthesis 30
tert-Butyl 3-(azidomethyl)-3-(hydroxymethyl)
pyrrolidine-1-carboxylate (44)
##STR00129##
[0706] Diol 43 (750 mg, 3.2 mmol) was dissolved in CH.sub.2Cl.sub.2
and pyridine (3.6 mL, 1:1). The mixture was cooled to 0.degree. C.
and treated with pTsCl (0.648 mg, 3.4 mmol) and DMAP (2 mg, 0.02
mmol). The mixture was stirred for 24 h at room temperature. The
mixture was concentrated under reduced pressure, dissolved in DMF
(15 ml), treated with NaN.sub.3 (1.85 mg, 22.4 mmol) and the
mixture was heated to 85.degree. C. for 24 h. The mixture was
re-cooled to room temperature, filtered and the filtrate was
extracted with EtOAc and washed with brine. The combined organic
layers were dried (Na.sub.2SO.sub.4) and concentrated under reduced
pressure. The title azide was obtained as a colorless oil (434 mg,
53%) after flash column chromatography (hexane:EtOAc gradient 9:1
to 7:3).
[0707] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 3.57 (d, J=3.7 Hz,
2H), 3.44 (m, 4H), 3.22 (td, J=18.1, 10.8 Hz, 2H), 2.17 (m, 1H),
1.78 (m, 1H), 1.45 (s, 9H); MS (Cl): m/z 279.2 (M+Na.sup.+).
Synthesis 31
tert-Butyl
3-(azidomethyl)-3-((methoxy-methoxy)methyl)pyrrolidine-1-carbox-
ylate (45)
##STR00130##
[0709] Azide 44 (650 mg, 2.54 mmol) was dissolved in
CH.sub.2Cl.sub.2 (8 mL) and treated with iso-Pr2NEt (1.8 mL, 10.33
mmol). The mixture was cooled to 0.degree. C., allowed to react
with MOMCl (0.6 mL, 7.9 mmol) and stirred at room temperature
overnight. The title azide was obtained (535 mg, 70%) after flash
column chromatography (hexane:EtOAc gradient 9:1 to 4:1).
[0710] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 4.62 (s, 2H), 3.43
(m, 6H), 3.36 (s, 3H), 3.22 (m, 2H), 1.79 (s, 2H), 1.45 (m, 9H);
.sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 154.5, 96.6, 79.5, 69.2,
55.4, 54.7, 51.8, 44.4, 30.9, 30.3, 28.5.
Synthesis 32
tert-Butyl
3-(aminomethyl)-3-((methoxy-methoxy)methyl)pyrrolidine-1-carbox-
ylate (46)
##STR00131##
[0712] Azide 45 (460 mg, 15.3 mmol) in MeOH (8 mL) was allowed to
react with Pd/C (10% wt., 50 mg) and stirred under an atmosphere of
hydrogen for 2 h. The mixture was filtered through a membrane
filter. The title amine was obtained as a colorless oil (400 mg,
95%), which was used without further purification.
[0713] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 4.61 (s, 2H), 3.45
(m, 4H), 3.36 (s, 3H), 3.20 (m, 2H), 2.76 (m, 2H), 1.75 (m, 2H),
1.45 (s, 9H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 154.5,
96.6, 79.5, 70.1, 55.4, 54.7, 51.8, 44.4, 30.9, 30.3, 28.5; MS
(ESI): m/z 275.2 (M+H.sup.+).
3.6-Synthesis of Pyrrolidine 51
##STR00132##
[0714] Synthesis 33
(2S,4R)-tert-Butyl
2-((tert-butyldimethyl-silyloxy)methyl)-4-hydroxypyrrolidine-1-carboxylat-
e (48)
##STR00133##
[0716] Imidazole (0.313 g, 4.6 mmol) and TBSCl (0.415 g, 2.76 mmol)
were added to a solution of alcohol 47 (0.50 g, 2.30 mmol) in
CH.sub.2Cl.sub.2 (5 mL) and the reaction mixture was stirred at
room temperature for 24 h. The reaction mixture was poured onto
water (5 mL), extracted with Et20 (5.times.5 mL) and the combined
organic layers were washed with brine, dried over MgSO.sub.4, and
concentrated under reduced pressure. Purification by flash column
chromatography (hexane:EtOAc gradient 2:1 to 1:1) gave 48 (0.480 g,
63%) as a clear oil. See, e.g., Vince et al., 1991.
[0717] R.sub.f=0.33 (hexane:EtOAc 1:1);
[.alpha.].sup.23.sub.D=-54.44 (c 1.15, CHCl.sub.3); IR (neat):
.nu..sub.max=3427, 1696, 1670, 1399, 1252, 1165, 1109 cm.sup.-1;
HRMS (ESI) calc. for C.sub.16H.sub.34NO.sub.4Si (M+H.sup.+):
332.2257, found: 332.2246.
Synthesis 34
(2S,4R)-tert-Butyl
2-((tert-butyldimethyl-silyloxy)methyl)-4-(diphenoxyphos-phoryloxy)pyrrol-
idine-1-carboxylate (49)
##STR00134##
[0719] DPPA (0.327 mL, 1.52 mmol) and DBU (0.228 mL, 1.52 mmol)
were added dropwise with stirring to a solution of alcohol 48
(0.420 g, 1.27 mmol) in THF (2.5 mL) at 0.degree. C. and the
mixture was allowed to warm to room temperature. After 24 h, the
reaction mixture was diluted with Et.sub.2O (5 mL), poured onto
saturated aqueous NaHCO.sub.3 (5 mL) and extracted with Et.sub.2O
(5.times.5 mL). The combined organic layers were washed with brine,
dried over MgSO.sub.4, and concentrated under reduced pressure.
Purification by flash column chromatography (hexane:EtOAc 4:1) gave
phosphate 49 (655 mg, 91%) as a clear oil.
[0720] R.sub.f=0.64 (hexane:EtOAc 1:1); IR (neat):
.nu..sub.max=1695, 1488, 1397, 1187, 1162 cm.sup.-1; HRMS (ESI)
calc. for C.sub.28H.sub.43NO.sub.7SiP (M+H.sup.+): 564.2546, found:
564.2563.
Synthesis 35
(2S,4S)-tert-Butyl-4-azido-2-((tert-butyl-dimethylsilyloxy)methyl)pyrrolid-
ine-1-carboxylate (50)
##STR00135##
[0722] A suspension of phosphate 49 (0.655 g, 1.16 mmol) and
NaN.sub.3 (0.9 g, 11.6 mmol) in DMF (2 mL) was heated to 80.degree.
C. for 3 days. The reaction mixture was diluted with Et.sub.2O (10
mL), poured onto water (10 mL) and extracted with Et.sub.2O
(6.times.5 mL). The combined organic layers were washed with water,
brine, dried over MgSO.sub.4 and concentrated under reduced
pressure. Purification by flash column chromatography (hexane:EtOAc
8:1) gave azide 50 (188 mg, 46%) as a clear oil. See, e.g.,
Moriarty et al., 2001.
[0723] R.sub.f=0.65 (hexane:EtOAc 4:1);
[.alpha.].sup.31.sub.D=-13.14 (c 1.11, CHCl.sub.3); IR (neat):
.nu..sub.max=2100, 1695, 1389, 1254, 1165 cm.sup.-1; .sup.13C NMR
(100 MHz, DMSO-d.sub.6, 353K) .delta. 153.0, 78.5, 62.7, 57.9,
57.0, 51.1, 31.7, 27.7, 25.3, 17.4, -5.8, -5.9; HRMS (ESI) calc.
for C.sub.16H.sub.32N.sub.4O.sub.3Si (M+H.sup.+): 357.2322, found:
357.2318.
Synthesis 36
(2S,4S)-tert-Butyl
4-amino-2-((tert-butyl-dimethylsilyloxy)methyl)pyrrolidine-1-carboxylate
(51)
##STR00136##
[0725] A solution of azide 50 (0.17 g, 0.477 mmol) in MeOH (2 mL)
was added with stirring to a suspension of Pd/C (10% wt., 25 mg) in
MeOH (1 mL) and the mixture was stirred for 2 h under an atmosphere
of hydrogen. The flask was purged with N.sub.2, the Pd/C filtered
off and the filtrate was concentrated under reduced pressure. The
crude product was filtered through a short plug of silica gel
(CHCl.sub.3:MeOH 9:1) to give amine 51 (128 mg, 81%) as a clear
oil. See, e.g., Moriarty et al., 2001.
[0726] R.sub.f=0.27 (CHCl.sub.3:MeOH 9:1);
[.alpha.].sup.23.sub.D=-27.1 (c 0.92, CHCl.sub.3); IR (neat):
.nu..sub.max=3211, 1694, 1474, 1364, 1385, 1252 cm.sup.-1; .sup.1H
NMR (400 MHz, DMSO-d.sub.6, 353K) .delta. 3.75-3.67 (m, 3H),
3.43-3.52 (m, 2H), 2.99 (dd, J=10.9, 7.1 Hz, 1H), 2.25-2.31 (m,
1H), 1.76-1.85 (m, 1H), 1.40 (s, 9H), 0.89 (s, 9H), 0.05 (s, 6H);
.sup.13C NMR (100 MHz, DMSO-d.sub.6, 353K) .delta.153.0, 78.3,
63.3, 57.1, 51.9, 48.0, 34.3, 27.8, 17.5, -5.8; HRMS (ESI) calc.
for C.sub.16H.sub.34N.sub.2O.sub.3Si (M+H.sup.+): 331.2417, found:
331.2413.
3.7-Synthesis of Piperidine 62
##STR00137## ##STR00138##
[0728] Synthesis 37
tert-Butyl 3,4-dihydroxypiperidine-1-carboxylate (53)
##STR00139##
[0730] Carbamate 52 (5.0 g, 27.3 mmol) was added to a solution of
potassium osmate (0.100 g, 0.271 mmol) and NMO (6.4 g, 54.6 mmol)
in THF and H.sub.2O (4:1; 50 mL). The mixture was stirred for 16 h
and a solution of sodium metabisulfite (30 mL) was added to quench
the excess oxidant. The aqueous layer was extracted with EtOAc
(5.times.30 mL). The combined organic layers were washed with
brine, dried over MgSO.sub.4 and concentrated under reduced
pressure. Purification by flash column chromatography (EtOAc) gave
diol 53 (5.23 g, 88%) as a clear oil. See, e.g., Ashton et al.,
2004.
[0731] R.sub.f=0.23 (EtOAc); IR (neat): .nu..sub.max=3355, 3253,
1665, 1423 cm.sup.-1; .sup.1H NMR (500 MHz, DMSO-d.sub.6, 353 K)
.delta. 4.24 (d, J=4.5 Hz, 1H), 4.14 (d, J=3.8 Hz, 1H), 3.69 (dq,
J=7.0, 3.4 Hz, 1H), 3.46 (dq, J=7.0, 3.4 Hz, 1H), 3.23-3.32 (m,
4H), 1.62-1.68 (m, 1H), 1.44-1.49 (m, 1H), 1.39 (s, 9H); .sup.13C
NMR (125 MHz, DMSO-d.sub.6, 353 K) .delta. 154.0, 78.0, 67.2, 59.2,
45.5, 29.3, 27.7, 20.2; HRMS (ESI) calc. for
C.sub.10H.sub.19NO.sub.4 (M+H.sup.+): 218.1392, found:
218.1394.
Synthesis 38
tert-Butyl 4-hydroxy-3-(tosyloxy)piperidine-1-carboxylate (55)
##STR00140##
[0733] pTsCl (4.46 g, 23.5 mmol) was added with stirring to a
solution of diol 53 (5.10 g, 23.5 mmol), Et.sub.3N (6.52 mL, 47.0
mmol), and DMAP (100 mg, 0.81 mmol) in CH.sub.2Cl.sub.2 (230 mL).
After 7 days, the reaction mixture was poured onto saturated
aqueous NH.sub.4Cl (100 mL) and extracted with EtOAc (5.times.50
mL). The combined organic layers were washed with brine, dried over
MgSO.sub.4 and concentrated under reduced pressure.
[0734] Purification by flash column chromatography (hexane:EtOAc
2:1) gave mono-4-toluenesulfonate 55 (1.30 g, 15%) as a white
solid.
[0735] R.sub.f=0.51 (EtOAc); IR (neat): .nu..sub.max=3416, 1660,
1437, 1352 cm.sup.-1; .sup.1H NMR (500 MHz, DMSO-d.sub.6, 353 K)
.delta. 7.80 (d, J=10.0 Hz, 2H), 7.43 (d, J=10.0 Hz, 2H), 4.43 (dt,
J=6.2, 2.9 Hz, 1H), 4.26 (brs, 1H), 3.99 (brs, 1H), 3.72-3.76 (m,
2H), 3.49-3.53 (m, 1H), 3.30 (dd, J=13.6, 3.0 Hz, 1H), 3.08 (ddd,
J=12.9, 7.6, 4.6 Hz, 1H), 2.41 (s, 3H), 1.54-1.58 (m, 2H), 1.37 (s,
9H); .sup.13C NMR (125 MHz, DMSO-d.sub.6, 353 K) .delta. 153.6,
143.9, 133.7, 129.3, 127.0, 78.6, 78.2, 65.6, 43.8, 29.2, 27.7,
20.5; HRMS (ESI) calc. for C.sub.17H.sub.25NO.sub.6S (M+Na.sup.+):
394.1300, found: 394.1295.
Synthesis 39
tent-Butyl 3-azido-4-hydroxypiperidine-1-carboxylate (57)
##STR00141##
[0737] NaN.sub.3 (1.11 g, 17.1 mmol) was added with stirring to a
solution of 4-toluenesulfonate 55 (1.26 g, 3.41 mmol) in DMF (10
mL). After 3 days, the reaction mixture was poured onto water (20
mL) and extracted with EtOAc (4.times.20 mL). The combined organic
layers were washed with brine, dried over MgSO.sub.4 and
concentrated under reduced pressure. Purification by flash column
chromatography (EtOAc) gave azide 57 (730 g, 88%) as a clear
oil.
[0738] R.sub.f=0.29 (hexane:EtOAc 1:1); IR (neat):
.nu..sub.max=3427, 2103, 1666, 1420, 1366 cm.sup.-1; .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 4.00 (dtd, J=13.7, 4.2, 1.8 Hz, 1H),
3.73 (t, J=6.2 Hz, 1H), 3.58 (brs, 1H), 3.28-3.32 (m, 1H),
2.83-2.87 (m, 1H), 2.64 (brs, 1H), 2.46 (t, J=6.2 Hz, 1H), 1.98
(dq, J=13.4, 3.8 Hz, 1H), 1.50 (s, 9H).
Synthesis 40
(3R,4R)-tert-Butyl
4-((R)-2-acetoxy-2-phenylacetoxy)-3-azidopiperidine-1-carboxylate
(58)
(3S,4S)-tent-Butyl
4-((R)-2-acetoxy-2-phenylacetoxy)-3-azidopiperidine-1-carboxylate
(59)
##STR00142##
[0740] (R)--O-acetoxy-mandelic acid (0.795 g, 3.84 mmol), DMAP (31
mg, 0.256 mmol) and EDCI (0.733 g, 3.84 mmol) were added to a
solution of azide 57 (0.620 g, 2.56 mmol) in CH.sub.2Cl.sub.2 (12
mL) and the reaction was stirred at ambient temperature. After 18 h
the reaction mixture was poured on water (20 mL) and extracted with
Et.sub.2O (4.times.25 mL). The combined organic layers were washed
with brine (20 mL), dried over MgSO.sub.4, and concentrated under
reduced pressure. The diastereoisomeric mixture was separated by
preperative HPLC to afford ester 58 (218 mg, 20%) and ester 59 (250
mg, 23%) as clear oils.
[0741] 58: R.sub.f=0.38 (hexane:EtOAc 4:1); IR (neat):
.nu..sub.max=2105, 1742, 1692, 1420, 1366 cm.sup.-1; .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 7.57-7.51 (m, 2H), 7.46-7.40 (m,
3H), 5.98 (s, 1H), 4.82 (td, J=7.9, 4.0 Hz, 1H), 3.90-3.65 (m, 2H),
3.35-3.00 (m, 3H), 2.16 (s, 3H), 1.81-1.73 (m, 1H), 1.40 (s, 9H),
1.32-1.21 (m, 1H); .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta.
170.4, 168.1, 154.0, 133.9, 129.7, 129.2, 128.1, 79.8, 74.5, 73.7,
72.8, 44.6, 28.3, 27.4, 20.7.
Synthesis 41
(3R,4R)-tert-Butyl 3-azido-4-hydroxy-piperidine-1-carboxylate
(60)
##STR00143##
[0743] LiOH (55 mg, 1.3 mmol) was added to a solution of ester 58
in THF, H.sub.2O and MeOH (1:1:1; 3 mL) and the mixture was stirred
for 2 h. The reaction mixture was poured onto water (5 mL) and
extracted with EtOAc (4.times.25 mL). The combined organic layers
were washed with brine, dried over MgSO.sub.4 and concentrated
under reduced pressure. Purification by flash column chromatography
(hexane:EtOAc 2:1) gave alcohol 60 as a clear oil (124 mg,
99%).
[0744] R.sub.f=0.29 (hexane:EtOAc 1:1); IR (neat):
.nu..sub.max=3427, 2103, 1666, 1420, 1366 cm.sup.-1; .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 4.00 (dtd, J=13.7, 4.2, 1.8 Hz, 1H),
3.73 (t, J=6.2 Hz, 1H), 3.58 (brs, 1H), 3.28-3.32 (m, 1H),
2.83-2.87 (m, 1H), 2.64 (brs, 1H), 2.46 (t, J=6.2 Hz, 1H), 1.98
(dq, J=13.4, 3.8 Hz, 1H), 1.50 (s, 9H); .sup.13C NMR (100 MHz,
CDCl.sub.3) .delta. 154.4, 80.5, 72.0, 63.3, 45.3, 41.2, 32.0,
28.3.
Synthesis 42
(3R,4R)-tert-Butyl
3-azido-4-(methoxy-methoxy)piperidine-1-carboxylate (61)
##STR00144##
[0746] iso-Pr.sub.2NEt (0.26 mL, 1.5 mmol) and MOMCl (0.075 mL, 1.0
mmol) were added with stirring to a solution of alcohol 60 (132 mg,
0.5 mmol) in CH.sub.2Cl.sub.2 (1 mL). After 18 h, additional
iso-Pr.sub.2NEt (0.26 mL, 1.5 mmol) and MOMCl (0.075 mL, 1.0 mmol)
were added.
[0747] After 24 h, saturated aqueous NaHCO.sub.3 (5 mL) was added,
the organic layer was separated and the aqueous layer was extracted
with EtOAc (3.times.10 mL). The combined organic layers were washed
with brine, dried over MgSO.sub.4, and concentrated under reduced
pressure. Purification by flash column chromatography (hexane:EtOAc
4:1) gave 61 (126 mg, 88%) as a clear oil.
[0748] R.sub.f=0.47 (hexane:EtOAc 2:1); [.alpha.].sup.25.sub.D=-9.7
(c 1.0, CHCl.sub.3); IR (neat): .nu..sub.max=2104, 1693, 1418,
1238, 1151 cm.sup.-1; .sup.1H NMR (500 MHz, CDCl.sub.3) .delta.
4.75 (d, J=7.0 Hz, 1H), 4.73 (d, J=7.0 Hz, 1H), 4.08 (brs, 1H),
3.84 (dtd, J=13.7, 4.6, 1.7 Hz, 1H), 3.54 (ddd, J=11.6, 6.7, 2.3
Hz, 1H), 3.36-3.42 (m, 4H), 2.93 (brs, 2H), 2.02 (dtd, J=13.4, 4.7,
3.2 Hz, 1H), 1.47-1.53 (m, 1H), 1.45 (s, 9H); .sup.13C NMR
(CDCl.sub.3, 125 MHz) .delta. 154.4, 95.5, 80.2, 61.0, 55.6, 45.4,
40.5, 30.1, 29.3, 28.4; HRMS (Cl) calc. for
C.sub.12H.sub.22N.sub.4O.sub.4 (M+H.sup.+): 287.1719, found:
287.1725.
Synthesis 43
(3R,4R)-tert-Butyl
3-amino-4-(methoxy-methoxy)piperidine-1-carboxylate (62)
##STR00145##
[0750] A solution of azide 61 (126 mg, 0.44 mmol) in MeOH (2 mL)
was added to a suspension of Pd/C (15% wt., 50 mg) in methanol (1
mL). The reaction mixture was stirred under an atmosphere of
hydrogen for 1 h, then filtered and concentrated under reduced
pressure. Purification by flash column chromatography
(CH.sub.2Cl.sub.2:MeOH 9:1) gave amine 62 (112 mg, 99%) as a clear
oil.
[0751] R.sub.f=0.25 (CH.sub.2Cl.sub.2:MeOH 9:1);
[.alpha.].sup.25.sub.D=+10.0 (c 1.0, CHCl.sub.3); IR (neat):
.nu..sub.max=3376, 1689, 1421, 1241, 1165 cm.sup.-1; .sup.1H NMR
(500 MHz, CDCl.sub.3) .delta. 4.76 (d, J=6.9 Hz, 1H), 4.69 (d,
J=6.9 Hz, 1H), 4.04 (s, 2H), 3.39 (s, 3H), 3.28-3.33 (m, 1H),
2.72-2.90 (m, 2H), 2.64 (t, J=11.7 Hz, 1H), 2.01 (ddd, J=9.5, 9.5,
5.0 Hz, 1H), 1.81 (s, 2H), 1.44-1.48 (m, 10H); .sup.13C NMR (100
MHz, CDCl.sub.3) .delta. 154.7, 95.7, 80.5, 79.8, 55.5, 52.4, 48.7,
42.1, 29.9, 28.4; HRMS (ESI) calc. for
Cu.sub.11H.sub.24N.sub.2O.sub.4 (M+H.sup.+): 261.1812, found:
261.1809.
4-Synthesis of the Target Compounds
4.1.1-General Procedure C Buchwald-Hartwig Coupling
[0752] The heteroaryl chloride (0.50 mmol), Pd.sub.2dba.sub.3 (23.0
mg, 5 mol %), rac-BINAP (47.0 mg, 15 mol %), and NaOtBu (72.0 mg,
0.75 mmol), were suspended in toluene (1.8 mL). After 5 min of
stirring, the pyrrolidine or piperidine (0.60 mmol) was added and
the mixture heated for 16 h at 95.degree. C. The reaction mixture
was cooled to room temperature, filtered through celite and washed
with EtOAc (10 mL) and poured onto brine (5 mL). The aqueous phase
was extracted with ethyl acetate (3.times.10 mL) and the combined
organic phases were dried over MgSO.sub.4 and concentrated under
reduced pressure. Purification by flash column chromatography
(hexane:EtOAc) gave the corresponding product. See, e.g., Hong et
al., 1997.
4.1.2-General Procedure D-Final Deprotection
[0753] The carbamate (0.15 mmol) was dissolved in MeOH and HCl
(generated by treatment of MeOH (5 mL) with acetyl chloride (2.5
mL) at room temperature for 45 min) (5 M, 8 mL) and stirred at room
temperature for 3 h. The solvent was removed under reduced pressure
and the residue was purified by flash column chromatography
(CH.sub.2Cl.sub.2:MeOH 9:1) to give the corresponding product.
4.2-Synthesis of PPDA-001
##STR00146##
[0754] Synthesis 44
(3R,4R)-tent-butyl
4-(((7-(benzyl(tert-butoxycarbonyl)amino)-3-iso-propylpyrazolo[1,5-a]pyri-
midin-5-Aamino)methyl)-3-(methoxymethoxy)piperidine-1-carboxylate
(63)
##STR00147##
[0756] Following general procedure C, chloride 14 (1.28 g, 3.21
mmol), Pd.sub.2dba.sub.3 (147 mg, 0.160 mmol), rac-BINAP (300 mg,
0.480 mmol), sodium tert-butoxide (370 mg, 3.85 mmol) and amine 38
(870 mg, 3.21 mmol) were allowed to react in toluene (2 mL).
Carbamate 63 was obtained as a pale yellow solid (1.48 g, 75%)
after flash column chromatography (hexane: EtOAc 1:5).
[0757] .sup.1H NMR .delta. (400 MHz, DMSO-d.sub.6, 363 K) .delta.
7.65 (s, 1H), 7.26 (m, 5H), 6.69 (brt, J=5.4 Hz, 1H), 6.01 (s, 1H),
4.85 (s, 2H), 4.65 (d, J=6.5 Hz, 1H), 4.60 (d, J=6.5 Hz, 1H), 4.04
(m, 1H), 3.73 (dt, J=13.1, 4.2 Hz, 1H), 3.63 (dt, J=13.6, 5.6 Hz,
1H), 3.31 (m, 1H), 3.29 (s, 3H), 3.04 (sept, J=6.9 Hz, 1H), 2.77
(dd, J=12.9, 8.8 Hz, 1H), 1.79 (m, 2H), 1.41 (s, 9H), 1.34 (s, 9H),
1.30 (d, J=6.9 Hz, 6H), 1.24 (m, 1H); .sup.13C NMR (100 MHz,
DMSO-d.sub.6, 363 K) .delta. 154.7, 153.4, 152.4, 145.4, 141.7,
140.0, 136.8, 127.5, 126.9, 126.5, 111.1, 96.7, 95.0, 80.5, 78.1,
74.0, 54.3, 50.7, 46.4, 41.8, 41.4, 40.4, 27.5, 27.1, 26.5, 22.8,
22.2.
Synthesis 45
(3R,4R)-4-(((7-(Benzylamino)-3-isopropylpyrazolo[1,5-a]pyrimidin-5-yl)amin-
o)methyl)piperidin-3-ol hydrochloride (PPDA-001)
##STR00148##
[0759] Following general procedure D, carbamates 63 (1.48 g, 2.41
mmol), was allowed to react with 5 M methanolic HCl. PPDA-001 was
obtained as a white solid (1.04 g, 99%) after flash column
chromatography (CH.sub.2Cl.sub.2:MeOH gradient 19:1 to 6:1).
[0760] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.90 (s, 1H), 7.45
(d, J=7.2 Hz, 2H), 7.39 (t, J=7.7 Hz, 2H), 7.32 (t, J=7.2 Hz, 1H),
5.48 (s, 1H), 4.75 (brs, 2H), 3.73 (dt, J=10.2, 4.5 Hz, 1H), 3.60
(dd, J=14.1, 3.9 Hz, 1H), 3.50 (brs, 1H), 3.46 (dd, J=2.2, 4.2 Hz,
1H), 3.37 (dt, J=12.8, 2.8 Hz, 1H), 3.12 (sept, J=6.8 Hz, 1H), 2.96
(td, J=12.7, 2.9 Hz, 1H), 2.81 (dd, J=11.4, 11.1 Hz, 1H), 2.05 (m,
1H), 1.86 (m, 1H), 1.65 (m, 1H), 1.33 (d, J=6.8 Hz, 6H); .sup.13C
NMR (100 MHz, CD.sub.3OD) .delta. 148.3, 142.2, 136.5, 128.6,
127.5, 127.0, 111.2, 65.3, 47.9, 45.3, 43.6, 43.0, 41.2, 24.5,
22.6, 22.2, 22.1.
4.3-Synthesis of PPDA-002
##STR00149##
[0761] Synthesis 46
(3aR,7 aR)-tert-Butyl
7a-((7-(benzyl(tert-butoxycarbonyl)amino)-3-iso-propyl-pyrazolo[1,5-a]pyr-
imidin-5-ylamino)methyl)-2,2-dimethyltetrahydro-[1,3]dioxolo[4,5-c]pyridin-
e-5(6H)-carboxylate (64)
##STR00150##
[0763] Following general procedure C, chloride 14 (89 mg, 0.22
mmol), Pd.sub.2dba.sub.3 (10 mg, 0.01 mmol), rac-BI NAP (19 mg,
0.03 mmol), sodium tert-butoxide (53 mg, 0.55 mmol) and amine 31
(70 mg, 0.24 mmol) were allowed to react in toluene (2 mL).
Carbamate 64 was obtained as a colorless oil (75 mg, 41%) after
flash column chromatography (hexane:Et.sub.2O gradient 7:3). The
title compound (not analytically pure) was directly used for the
next step. .sub.Synthesis 47
(3R,4R)-4-((7-(Benzylamino)-3-iso-propyl-pyrazolo[1,5-a]pyrimidin-5-ylamin-
o) methyl)piperidine-3,4-diol hydrochloride (PPDA-002)
##STR00151##
[0765] Following general procedure D, carbamate 64 (50 mg, 0.077
mmol), was allowed to react with 5 M methanolic HCl. PPDA-002 was
obtained as a white solid (31 mg, 98%) after flash column
chromatography (CH.sub.2Cl.sub.2:MeOH 4:1).
[0766] R.sub.f=0.20 (CH.sub.2Cl.sub.2:MeOH 4:1); .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 7.90 (s, 1H), 7.43-7.29 (m, 5H), 5.43
(s, 1H), 4.72 (s, 2H), 3.66 (td, J=10.3, 4.4 Hz, 1H), 3.59-3.54 (m,
1H), 3.48-3.34 (m, 3H), 3.07 (sept, J=6.9 Hz, 1H), 2.93 (td,
J=12.6, 2.9 Hz, 1H), 2.77 (t, J=11.3 Hz, 1H), 2.05-2.00 (m, 1H),
1.86-1.78 (m, 1H), 1.63-1.53 (m, 1H), 1.31 (d, J=6.9 Hz, 6H);
.sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 156.6, 146.8, 145.3,
140.8, 136.9, 129.0, 128.0, 127.3, 112.9, 73.1, 71.5, 55.4, 53.5,
50.3, 46.1, 29.8, 23.9, 23.4.
4.4-Synthesis of PPDA-003
##STR00152##
[0767] Synthesis 48
(3R,4R)-tent-Butyl 4-((7-(tert-butoxycarbonyl
(pyridin-4-ylmethyl)amino)-3-iso-propyl
pyrazolo[1,5-a]pyrimidin-5-ylamino)methyl)-3-(methoxymethoxy)piperidine-1-
-carboxylate (65)
##STR00153##
[0769] Following general procedure C, chloride 15 (91 mg, 0.22
mmol), Pd.sub.2dba.sub.3 (15 mg, 0.016 mmol), rac-BINAP (20 mg,
0.033 mmol), sodium tert-butoxide (32 mg, 0.33 mmol) and amine 38
(75 mg, 0.273 mmol) were allowed to react in toluene (1 mL).
Carbamate 65 was obtained as a pale yellow solid (94 mg, 66%) after
flash column chromatography (EtOAc).
[0770] R.sub.f=0.30 (EtOAc); IR (neat): .nu..sub.max=1692, 1643,
1523, 1154 cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
8.51 (d, J=6.0 Hz, 2H), 7.72 (s, 1H), 7.22 (d, J=6.0 Hz, 2H), 5.82
(s, 1H), 5.39 (t, J=5.9 Hz, 1H), 4.92 (s, 2H), 4.73 (d, J=6.8 Hz,
1H), 4.62 (d, J=6.8 Hz, 1H), 4.30 (brs, 1H), 4.02-3.90 (m, 1H),
3.69-3.63 (m, 1H), 3.49-3.40 (m, 1H), 3.37 (s, 3H), 3.36-3.29 (m,
1H), 3.10 (sept, J=6.9 Hz, 1H), 2.65 (t, J=12.8 Hz, 1H), 2.54 (s,
1H), 1.80-1.67 (m, 2H), 1.43 (s, 9H), 1.37 (s, 9H), 1.31 (d, J=6.9
Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 154.8, 154.5,
153.4, 149.8, 149.4, 147.0, 146.3, 142.5, 141.7, 128.5, 122.4,
113.4, 97.0, 96.1, 82.7, 79.8, 76.0, 60.4, 55.9, 50.7, 43.3, 42.3,
28.3, 28.0, 23.8, 23.1.
Synthesis 49
(3R,4R)-4-((3-iso-Propyl-7-(pyridin-4-ylmethylamino)pyrazolo[1,5-a]pyrimid-
in-5-ylamino)methyl)piperidin-3-ol hydrochloride (PPDA-003)
##STR00154##
[0772] Following general procedure D, carbamate 65 (75 mg, 0.117
mmol), was allowed to react with 5 M methanolic HCl. PPDA-004 was
obtained as a white solid (23 mg, 46%) after flash column
chromatography (CH.sub.2Cl.sub.2:MeOH gradient 19:1 to 9:1).
[0773] R.sub.f=0.20 (CHCl.sub.3:MeOH 5:1); IR (neat):
.nu..sub.max=3278, 1717, 1643, 1584, 1156 cm.sup.-1; .sup.1H NMR
(500 MHz, CD.sub.3OD) .delta. 8.85 (d, J=6.2 Hz, 2H), 8.12 (d,
J=6.1 Hz, 2H), 5.15 (s, 2H), 3.68 (td, J=10.1, 4.4 Hz, 1H), 3.58
(d, J=11.5 Hz, 1H), 3.53-3.42 (m, 1H), 3.40-3.32 (m, 2H), 3.16-3.07
(m, 1H), 2.96 (t, J=10.5 Hz, 1H), 2.76 (t, J=11.3 Hz, 1H), 2.66 (s,
3H), 2.08 (d, J=14.5 Hz, 1H), 1.85 (brs, 1H), 1.70-1.59 (m, 1H),
1.32 (d, J=6.9 Hz, 6H); .sup.13C NMR (100 MHz, CD.sub.3OD, 353 K)
.delta. 160.3, 155.2, 151.1, 144.1, 142.9, 135.0, 126.6, 112.7,
66.7, 45.9, 45.3, 44.3, 42.4, 40.5, 25.9, 24.0, 23.5.
4.5-Synthesis of PPDA-007
##STR00155##
[0774] Synthesis 50
(3R,4R)-tent-Butyl 4-((7-(tert-butoxycarbonyl
(4-(4-methylpiperazin-1-yl)phenyl)amino)-3-iso-propylpyrazolo[1,5-a]pyrim-
idin-5-ylamino)methyl)-3-(methoxymethoxy) piperidine-1-carboxylate
(66)
##STR00156##
[0776] Following general procedure C, chloride 16 (100 mg, 0.20
mmol), Pd.sub.2dba.sub.3 (13 mg, 0.014 mmol), rac-BINAP (17 mg,
0.028 mmol), sodium tert-butoxide (29 mg, 0.30 mmol) and amine 38
(55 mg, 0.20 mmol) were allowed to react in toluene (2 mL).
Carbamate 66 was obtained as a white solid (69 mg, 48%) after flash
column chromatography (CH.sub.2Cl.sub.2:MeOH 20:1).
[0777] R.sub.f=0.46 (20:1 CH.sub.2Cl.sub.2/MeOH);
[.alpha.].sub.D.sup.23+20.3 (c 1.0, CHCl.sub.3); IR (neat):
.nu..sub.max=3370, 1698, 1643, 1515, 1157 cm.sup.-1; HRMS (ESI)
Calcd. for C.sub.38H.sub.58N.sub.8O.sub.6 [M+H].sup.+, 723.4558,
found 723.4548.
Synthesis 51
(3R,4R)-4-((3-iso-Propyl-7-(4-(4-methylpiperazin-1-yl)phenylamino)pyrazolo-
[1,5-a]pyrimidin-5-ylamino)methyl)piperidin-3-ol hydrochloride
(PPDA-007)
##STR00157##
[0779] Following general procedure D, carbamate 66 (40 mg, 0.055
mmol), was allowed to react with 5 M methanolic HCl. PPDA-008 was
obtained as a white solid (22 mg, 80%) after HPLC
(water:acetonitrile gradient 95:5 to 40:60).
[0780] IR (neat): .nu..sub.max=3246, 2474, 1659, 1575 cm.sup.-1;
.sup.13C NMR (CD.sub.3OD, 125 MHz) .delta. 150.4, 148.8, 143.9,
137.9, 135.1, 130.0, 128.9, 127.9, 118.9, 115.7, 103.9, 66.7, 57.0,
54.6, 46.6, 44.9, 44.4, 43.6, 42.6, 40.4, 34.6, 27.4, 26.1, 25.7,
23.8, 23.6.
4.6-Synthesis of PPDA-009
##STR00158##
[0781] Synthesis 52
(3R,4R)-tent-Butyl
4-(((7-(1-(tert-butoxycarbonyl)piperidine-4-carboxamido)-3-iso-propylpyra-
zolo[1,5-a]pyrimidin-5-yl)amino)methyl)-3-(methoxymethoxy)
piperidine-1-carboxylate (67)
##STR00159##
[0783] Following general procedure C, chloride 12 (230 mg, 0.546
mmol), Pd.sub.2dba.sub.3 (50 mg, 0.054 mmol), rac-BINAP (50 mg,
0.10 mmol), sodium tert-butoxide (61 mg, 0.82 mmol) and amine 38
(171 mg, 0.624 mmol) were allowed to react in toluene (2 mL).
Carbamate 67 was obtained as a pale yellow solid (220 mg, 61%)
after flash column chromatography (hexane: EtOAc 7:3).
[0784] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.14 (s, 1H), 7.66
(s, 1H), 6.93 (s, 1H), 5.25 (brs, 1H), 4.82 (d, J=6.8 Hz, 1H), 4.69
(d, J=6.8 Hz, 1H), 4.23 (brs, 3H), 4.04 (brs, 1H), 3.75-3.64 (m,
1H), 3.64-3.51 (m, 1H), 3.49-3.33 (m, 4 H), 3.13 (quin, J=6.8 Hz,
1H), 2.92-2.78 (m, 2H), 2.78-2.51 (m, 3H), 1.98 (d, J=11.7 Hz, 2H),
1.87-1.69 (m, 4 H), 1.52-1.46 (m, 18 H), 1.35 (d, J=6.8 Hz,
6H).
Synthesis 53
N-(5-((((3R,4R)-3-Hydroxypiperidin-4-yl)methyl)amino)-3-iso-propylpyrazolo-
[1,5-a]pyrimidin-7-yl)piperidine-4-carboxamide hydrochloride
(PPDA-009)
##STR00160##
[0786] Following general procedure D, carbamate 67 (100 mg, 0.15
mmol), was allowed to react with 5 M methanolic HCl. PPDA-010 was
obtained as a white solid (64 mg, 90%) after flash column
chromatography (CH.sub.2Cl2:MeOH gradient 10:0 to 85:15).
[0787] IR (neat): .nu..sub.max=3284, 1730, 1639, 1584 cm.sup.-1;
.sup.1H NMR (400MHz ,MeOD-d.sub.4) .delta. 7.98 (brs, 1H), 7.39
(brs, 1H), 3.73 (d, J=8.3 Hz, 2H), 3.57-3.35 (m, 4 H), 3.26-2.97
(m, 5 H), 2.86 (t, J=10.8 Hz, 1H), 2.26-2.16 (m, 3H), 2.13 (brs,
1H), 2.07-1.91 (m, 3H), 1.73 (brs, 1H), 1.33 (d, J=6.4 Hz, 6H).
4.7-Synthesis of PPDA-010
##STR00161##
[0788] Synthesis 54
tert-Butyl 4-(((7-((tert-butoxycarbonyl)
(pyridin-4-ylmethyl)amino)-3-iso-propylpyrazolo[1,5-a]pyrimidin-5-Aamino)-
methyl)-4-((triethylsilyl)oxy)piperidine-1-carboxylate (68)
##STR00162##
[0790] Following general procedure C, chloride 15 (1.182 g, 2.94
mmol), Pd.sub.2dba.sub.3 (119 mg, 0.13 mmol), rac-BI NAP (249 mg,
0.40 mmol), sodium tert-butoxide (385 mg, 4.01 mmol) and 40 (920
mg, 2.67 mmol) were allowed to react in toluene (2 mL). Carbamate
68 was obtained as a pale yellow solid (531 mg, 28%) after flash
column chromatography (hexane:EtOAc 1:5).
R.sub.f=0.5 (EtOAc); HRMS (ESI) calc. for
C.sub.37H.sub.60N.sub.7O.sub.5Si (M+H.sup.+): 710.4425, found:
710.4456.
Synthesis 55
4-(((3-iso-Propyl-7-((pyridin-4-ylmethyl)amino)pyrazolo[1,5-a]pyrimidin-5--
yl)amino)methyl)piperidin-4-ol hydrochloride (PPDA-010)
##STR00163##
[0792] Following general procedure D, carbamate 68 (500 mg, 0.70
mmol), was allowed to react with 5 M methanolic HCl. PPDA-011 was
obtained as a white solid (195 mg, 70%) after flash column
chromatography (CH.sub.2Cl.sub.2:MeOH gradient 19:1 to 6:1).
[0793] R.sub.f=0.4 (CHCl.sub.3:MeOH 9:1); IR (neat):
.nu..sub.max=3321, 1728, 1660, 1584, 1460, 1384, 1290, 1272, 1123
cm.sup.-1; .sup.1H NMR (CD.sub.3OD, 500 MHz) .delta. 8.80 (2H, d,
J=10.0 Hz), 8.05 (2H, d, J=10.0 Hz), 7.93 (1H, s), 7.72-7.70 (1H,
m), 7.62-7.61 (1H, m), 5.15 (2H, br s), 4.20 (1H, m), 3.51 (2H, s),
3.27-3.24 (4H, m), 3.11 (1H, m), 1.88 (5H, m), 1.33 (6H, d, J=10.0
Hz); HRMS (ESI) calc. for C.sub.21H.sub.30N.sub.7O (M+H.sup.+):
396.2512, found: 396.2504.
4.8-Synthesis of PPDA-015
##STR00164##
[0794] Synthesis 56
tert-Butyl-5-((3S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethoxy)pyrrolidin-
-3-ylamino)-3-iso-propylpyrazolo[1,5-a]pyrimidin-7-ylbenzylcarbamate
(69)
##STR00165##
[0796] Following general procedure C, chloride 14 (110 mg, 0.28
mmol), Pd.sub.2dba.sub.3 (13 mg, 0.014 mmol), rac-BINAP (427 mg,
0.042 mmol), sodium tert-butoxide (36 mg, 0.37 mmol) and 22 (89 mg,
0.36 mmol) were allowed to react in toluene (3 mL). Carbamate 69
was obtained as an orange oil (145 mg, 85%) after flash column
chromatography (hexane:Et.sub.2O 7:3).
[0797] R.sub.f=0.36 (Et.sub.2O:hexane:30% aqueous ammonia
71.75:28:0.25); [.alpha.].sup.20.sub.D=+33 (c 0.58,
CH.sub.2Cl.sub.2); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.76
(s, 1H), 7.23-7.30 (m, 5H), 5.76 (s, 1H), 5.29 (d, J=7.2 Hz, 0.6H
rotamers), 5.24 (d, J=7.2 Hz, 0.4H rotamers), 4.94 (br s, 2H), 4.67
(t, J=10.4 Hz, 1H), 4.55-4.63 (m, 2H), 4.27 (br s, 1H), 3.83-3.91
(m, 1H), 3.51-3.63 (m, 2H), 3.28 (s, 1.7H rotamers), 3.27 (s, 1.3H
rotamers), 3.12-3.23 (m, 2H), 1.47 (s, 3.7H rotamers), 1.45 (s,
5.3H rotamers), 1.40 (br s, 9H), 1.33 (d, J=6.8 Hz, 6H); .sup.13C
NMR (100 MHz, CDCl.sub.3) .delta. 154.6, 153.8, 146.1, 142.9,
141.6, 137.7, 128.5, 127.9, 127.5, 113.8, 97.5, 96.0, 82.2, 79.6,
77.4, 75.7, 55.7, 52.5, 51.4, 50.5, 48.8, 28.5, 28.1, 23.9, 23.7;
HRMS (ESI) calc. for C.sub.32H.sub.46N.sub.56O.sub.6 (M+H.sup.+):
611.3557, found 611.3543.
Synthesis 57
(3S,4S)-4-(7-(Benzylamino)-3-iso-propyl-pyrazolo[1,5-a]pyrimidin-5-ylamino-
)-pyrrolidin-3-ol hydrochloride (PPDA-015)
##STR00166##
[0799] Following general procedure D, carbamate 69 (140 mg, 0.23
mmol), was allowed to react with 5M methanolic HCl. PPDA-016 was
obtained as a pale orange solid (80 mg, 95%) after flash column
chromatography (CH.sub.2Cl.sub.2: MeOH 4:1).
[0800] R.sub.f=0.32 (CH.sub.2Cl.sub.2:MeOH:30% aqueous ammonia
80:19.5:0.5); [.alpha.].sup.20.sub.D=+12 (c 1.0,
[0801] MeOH); m.p.=102.degree. C.; .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.65 (s, 1H), 7.30 (brs, 5H), 6.59 (brs, 1H),
5.58 (d, J=4.8 Hz, 1H), 5.1 (s, 1H), 4.62 (brs, 2H), 4.33-4.37 (m,
3H), 4.26-4.31 (m, 1H), 3.33 (dd, J=11.2, 8.0 Hz, 1H), 3.15 (dd,
J=12.0, 4.0 Hz, 1H), 3.03-3.11 (m, 2H), 2.85 (dd, J=10.2, 7.2 Hz,
1H), 1.30 (d, J=6.8 Hz, 6H); .sup.13C NMR (100 MHz, CDCl.sub.3)
.delta. 156.6, 146.8, 145.3, 140.8, 136.9, 129.0, 128.0, 127.3,
112.9, 73.1, 71.5, 55.4, 53.5, 50.3, 46.1, 29.8, 23.9, 23.4; HRMS
(ESI) calc. for C.sub.20H.sub.26N.sub.6O (M+H.sup.+): 367.2246,
found 367.2239; Anal. calc. for C.sub.20H.sub.27ClN.sub.6O: C
65.55, H 7.15, N 22.93, found: C 65.54, H 7.09, N 22.87.
4.9-Synthesis of PPDA-018
##STR00167##
[0802] Synthesis 58
tert-Butyl 3-((7-(benzyl(tert-butoxycarbonyl)
amino)-3-iso-propylpyrazolo[1,5-a]pyrimidin-5-ylamino)methyl)-3-((methoxy-
methoxy) methyl)pyrrolidine-1-carboxylate (70)
##STR00168##
[0804] Following general procedure C, chloride 14 (879 mg, 2.2
mmol), Pd.sub.2dba.sub.3 (133 mg, 0.14 mmol) rac-BINAP (220 mg,
0.35 mmol), sodium tert-butoxide (220 mg, 2.3 mmol) and amine 46
(400 mg, 1.5 mmol) were allowed to react in toluene (15 mL).
Carbamate 70 was obtained as a pale yellow solid (600 mg, 65%)
after flash column chromatography (hexane:EtOAc gradient 19:1 to
7:3).
[0805] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.75 (s, 1H), 7.30
(m, 5H), 7.22 (m, 1H), 5.66 (m, 1H), 5.03 (m, 3H), 4.60 (s, 2H),
3.50-3.25 (m, 10H), 3.12 (td, J=13.8, 6.9Hz, 1H), 1.76 (m, 2H),
1.45 (d, J=9.3 Hz, 9H), 1.40 (s, 9H), 1.32 (d, J=6.8 Hz, 6H); MS
(ESI): m/z 639.4 (M+H.sup.+).
Synthesis 59
(3-((7-(Benzylamino)-3-iso-propylpyrazolo
[1,5-a]pyrimidin-5ylamino)methyppyrrolidin-3-yl)methanol
hydrochloride (PPDA-018)
##STR00169##
[0807] Following general procedure D, carbamate 70 (600 mg, 10.94
mmol), was allowed to react with 5 M methanolic HCl. PPDA-019 was
obtained as a white solid (320 mg, 80%) after flash column
chromatography (CH.sub.2Cl.sub.2:MeOH gradient 19:1 to 9:1).
[0808] IR (neat): .nu..sub.max=3274, 1663, 1577 cm.sup.-1; .sup.1H
NMR (400 MHz, CD.sub.3OD) .delta. 7.68 (s, 1H), 7.33 (m, 5H), 5.20
(s, 1H), 4.55 (s, 2H), 3.44 (m, 6H), 3.17 (q, J=12.2 Hz, 2H), 3.03
(m, 1H), 1.92 (m, 2H), 1.29 (dd, J=6.9, 3.9 Hz, 6H); .sup.13C NMR
(100 MHz, CD.sub.3OD) .delta. 148.5, 141.7, 139.1, 129.8, 128.6,
128.1, 113.6, 73.8, 64.8, 51.9, 50.9, 46.6, 46.2, 44.9, 31.6, 24.7,
23.9, 23.8; HRMS (ESI) calc. for C.sub.22H.sub.30N.sub.6O
(M+H.sup.+): 395.2481, found: 395.2534.
4.10-Synthesis of PPDA-022
##STR00170##
[0809] Synthesis 60
(3R,4R)-tent-Butyl
3-(7-(benzyl(tert-butoxy-carbonyl)amino)-3-iso-propylpyrazolo[1,5-a]pyrim-
idin-5-ylamino)-4 (methoxymethoxy) piperidine-1-carboxylate
(71)
##STR00171##
[0811] Following general procedure C, chloride 14 (72 mg, 0.18
mmol), Pd.sub.2dba.sub.3 (8 mg, 0.009 mmol), rac-BI NAP (12 mg,
0.018 mmol), sodium tert-butoxide (26 mg, 0.27 mmol) and amine 62
(47 mg, 0.18 mmol) were allowed to react in toluene (1 mL).
Carbamate 71 was obtained as a pale yellow oil (32 mg, 30%) after
flash column chromatography (hexane: EtOAc 6:1).
[0812] R.sub.f=0.3 (hexane: EtOAc 2:1); [.alpha.].sup.25.sub.D=+1.0
(c 1.11, CHCl.sub.3); IR (neat): .nu..sub.max=3347, 1719, 1670,
1640, 1366, 1153 cm.sup.-1; HRMS (ESI) calc. for
C.sub.33H.sub.48N.sub.6O.sub.6 (M+H.sup.+): 625.3714, found:
625.3708.
Synthesis 61
(3R,4R)-3-(7-(Benzylamino)-3-iso-propyl-pyrazolo[1,5-a]pyrimidin-5-ylamino-
) piperidin-4-ol (PPDA-022)
##STR00172##
[0814] Following general procedure D, carbamate 71 (32 mg, 0.05
mmol), was treated with 5 M methanolic HCl. PPDA-023 was obtained
as a white solid (16.2 mg, 76%) after flash column chromatography
(CH.sub.2Cl.sub.2:MeOH 9:1).
[0815] R.sub.f=0.13 (CHCl.sub.3:MeOH 5:1); IR (neat):
.nu..sub.max=3294, 1626, 1569, 1450 cm.sup.-1; .sup.1H NMR (500
MHz, CD.sub.3OD) .delta. 7.74 (s, 1H), 7.39 (d, J=7.6 Hz, 2H), 7.33
(tt, J=7.7, 1.5 Hz, 2H), 7.24-7.26 (m, 1H), 4.61 (s, 2H), 4.09 (dt,
J=10.2, 5.0 Hz, 1H), 3.84 (td, J=9.2, 3.8 Hz, 1H), 3.57-3.61 (m,
2H), 3.40 (dt, J=12.8, 4.5 Hz, 1H), 3.09 (ddt, J=13.5, 11.1, 3.0
Hz, 3H), 2.21-2.27 (m, 1H), 1.83 (dd, J=18.7, 7.9 Hz, 1H), 1.31 (d,
J=6Hz, 3H), 1.29 (d, J=6 Hz, 3H); .sup.13C NMR (125 MHz,
CD.sub.3OD) .delta. 156.9, 149.2, 144.1, 138.6, 129.8, 129.0,
128.7, 113.5, 69.0, 67.6, 53.4, 47.5, 46.6, 42.9, 30.3, 28.3, 24.6,
23.7; HRMS (ESI) calc. for C.sub.21H.sub.28N.sub.6O (M+H.sup.+):
381.2403, found: 381.2400.
4.11-Synthesis of PPDA-026
##STR00173##
[0816] Synthesis 62
(2S,4S)-tert-Butyl
4-(7-(benzyl(tert-butoxy-carbonyl)amino)-3-iso-propyl
pyrazolo[1,5-a]pyrimidin-5-ylamino)-2-((tert-butyl
dimethylsilyloxy)methyl) pyrrolidine-1-carboxylate (72)
##STR00174##
[0818] Following general procedure C, chloride 14 (45.1 mg, 0.112
mmol), Pd.sub.2dba.sub.3 (5 mg, 0.0056 mmol), rac-BINAP (8 mg,
0.011 mmol), sodium tert-butoxide (16.1 mg, 0.168 mmol) and amine
51 (44.7 mg, 0.135 mmol) were allowed to react in toluene (2 mL).
Carbamate 72 was obtained as a pale yellow solid (45.5 mg, 59%)
after flash column chromatography (hexane: EtOAc 4:1).
[0819] R.sub.f=0.55 (hexane:EtOAc 2:1);
[.alpha.].sup.24.sub.D=-72.4 (c 1.7, CHCl.sub.3); IR (neat):
.nu..sub.max=3343, 1692, 1641, 1518, 1390, 1366, 1252, 1157
cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.77 (s, 1H),
7.24-7.36 (m, 5H), 5.96-5.98 (m, 1H), 5.70 (s, 2H), 4.64 (s, 1H),
4.23-4.26 (m, 1H), 3.78-4.01 (m, 2H), 3.60-3.67 (m, 1H), 3.80-3.27
(m, 1H), 3.14 (sept, J=8.0 Hz, 1H), 2.43-2.51 (m, 1H), 1.88 (d,
J=16.0 Hz, 1H), 1.47 (s, 9H), 1.43 (s, 9H), 1.36 (d, J=8.0 Hz, 6H),
0.9 (s, 9H), 0.09 (s, 3H), 0.06 (s, 3H); HRMS (ESI) calc. for
C.sub.37H.sub.58N.sub.6O.sub.5Si (M+H.sup.+): 695.4316, found:
695.4330.
Synthesis 63
((2S,4S)-4-(7-(Benzylamino)-3-iso-propyl
pyrazolo[1,5-a]pyrimidin-5-ylamino) pyrrolidin-2-yl)methanol
hydrochloride (PPDA-026)
##STR00175##
[0821] Following general procedure D, carbamate 72 (37.5 mg, 0.054
mmol), was allowed to react with 5M methanolic HCl. PPDA-027 was
obtained as a yellow solid (20.9 mg, 85%) after flash column
chromatography (CH.sub.2Cl.sub.2:MeOH gradient 9:1 to 5:1).
[0822] R.sub.f=0.22 (CHCl.sub.3: MeOH 9:1);
[.alpha.].sup.27.sub.D=-12.6 (c 0.80, MeOH); IR (neat):
.nu..sub.max=3235, 1654, 1576 cm.sup.-1; HRMS (ESI) calc. for
C.sub.21H.sub.28N.sub.6O (M+H.sup.+): 381.2403, found:
381.2398.
5-Optimised Synthesis of PPDA-001
##STR00176## ##STR00177##
[0823] Synthesis 64
1-tert-butyl 4-ethyl 3-oxopiperidine-1,4-dicarboxylate (74)
##STR00178##
[0825] A mixture of ethyl 1-benzyl-3-oxo-4-piperidinecarboxylate
hydrochloride (10.0 g, 33.58 mmol), Pd/C (10% wt., 1.0 g),
Boc.sub.2O (14.64 g, 67.16 mmol), Na.sub.2CO.sub.3 (3.56 g, 33.58
mmol) and EtOH (100 mL) was loaded into a Parr autoclave. Hydrogen
was introduced (38 bar), and the mixture was stirred at 50.degree.
C. for 48 h. After the autoclave was cooled to 25.degree. C. the
hydrogen pressure was released, the catalyst was removed by
filtration, and the mixture was concentrated under reduced pressure
to give a yellow oil. Purification by column chromatography
(hexane:EtOAc 2:1) gave the title compound (9.10 g, 100%) as a
clear oil.
[0826] R.sub.f=0.70 (hexane: EtOAc 2:1); .sup.1H NMR (CDCl.sub.3,
400 MHz) .delta. 12.10 (s, 1H, enol form --OH), 4.25 (q, J=7.1 Hz,
2H), 4.04 (s, 2H), 3.50 (t, J=5.8 Hz, 2H), 2.33 (t, J=5.8 Hz, 2H),
1.48 (s, 9H), 1.32 (t, J=7.1 Hz, 3H); HRMS (Cl) calc. for
C.sub.13H.sub.21NO.sub.5 (M+NH.sub.4).sup.+ 289.1763, found:
289.1759.
Synthesis 65
(3R,4S)-1-tert-butyl 4-ethyl 3-hydroxypiperidine-1,4-dicarboxylate
(75)
##STR00179##
[0828] A solution of 1-tert-butyl 4-ethyl
3-oxopiperidine-1,4-dicarboxylate 74 (4.46 g, 16.47 mmol) and
CH.sub.2Cl.sub.2 (20 mL) was added to a 450 mL glass liner
containing a stirring bar and degassed by bubbling nitrogen for 30
min. [RuCl(p-cymene)(S)-T-BINAP]Cl complex (0.324 g, 0.33 mmol, 2
mol %) was added and the liner was loaded into a Parr autoclave.
Hydrogen was introduced (10 bar), and the mixture was heated to
50.degree. C. for 48 h. After the autoclave was cooled to
25.degree. C. the hydrogen pressure was released, and the mixture
was concentrated to give a red oil. Purification by column
chromatography (hexane:EtOAc gradient 6:1 to 3:1) gave the title
compound (3.47 g, 77%) as a clear oil. The enantiomeric ratio was
determined to be 97:3 by HPLC analysis after converting an aliquot
of the product to the (R)-acetylmandelic ester.
[0829] R.sub.1=0.25 (hexane:EtOAc 2:1); [.alpha.]D.sub.22+14.2 (c
1.41, CHCl.sub.3); .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
4.27-4.22 (m, 1H), 4.21 (q, J=7.1 Hz, 2H), 3.83 (td, J=9.9, 4.8 Hz,
1H), 3.13 (brs, 1H), 2.71 (brs, 1H), 2.61 (dd, J=13.0, 10.3 Hz,
1H), 2.38 (ddd, J=12.3, 9.5, 4.1 Hz, 1H), 2.05-1.97 (m, 1H),
1.64-1.53 (m, 1H), 1.46 (s, 9H), 1.29 (t, J=7.1 Hz, 3H); .sup.13C
NMR (CDCl.sub.3, 100 MHz) .delta. 173.9, 154.5, 80.0, 67.4, 49.2,
48.9, 42.7, 28.4, 26.7, 14.1; HRMS (ESI) calc. for
C.sub.13H.sub.23NO.sub.5 (M+Na).sup.+ 296.1474, found:
296.1486.
Synthesis 66
(3R,4S)-1-tert-butyl 4-ethyl
3-(methoxymethoxy)piperidine-1,4-dicarboxylate (76)
##STR00180##
[0831] To a solution of (3R,4S)-1-tert-butyl 4-ethyl
3-hydroxypiperidine-1,4-dicarboxylate (4.10 g, 15.0 mmol) in
CH.sub.2Cl.sub.2 (75 mL) was added DIPEA (10.18 mL, 60.0 mmol) and
MOMCl (3.42 mL, 45.0 mmol). After 24 h, NaHCO.sub.3 (50 mL) was
added the organic layer was separated and the aqueous layer was
extracted with EtOAc (4.times.30 mL). The combined organic layers
were washed with NH.sub.4Cl (30 mL), brine (30 mL), dried over
MgSO.sub.4, and concentrated under reduced pressure to give a
yellow oil. Purification by column chromatography (hexane:EtOAc
4:1) gave the title compound as a clear oil (3.56 g, 75%).
[0832] R.sub.f=0.65 (hexane:EtOAc 1:1); [.alpha.].sub.D.sup.22+2.3
(c 1.12, CHCl.sub.3); .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
4.68 (s, 2H), 4.17 (q, J=7.2 Hz, 2H), 4.00-3.96 (m, 1H), 3.80-3.76
(m, J=9.6, 4.7 Hz, 2H), 3.35 (s, 3H), 2.78 (ddd, J=13.8, 11.8, 3.0
Hz, 1H), 2.70 (brs, 1H), 2.50 (ddd, J=11.4, 9.3, 4.1 Hz, 1H), 1.91
(dq, J=13.5, 3.6 Hz, 1H), 1.71-1.62 (m, 1H), 1.46 (s, 9H), 1.27 (t,
J=7.1 Hz, 3H); .sup.13C NMR (CDCl.sub.3, 100 MHz) .delta. 173.4,
154.5, 96.3, 79.9, 73.3, 60.7, 55.6, 48.5, 47.1, 42.4, 28.3, 27.4,
14.2; HRMS (ESI) calc. for C.sub.15H.sub.27NO.sub.6 (M+H).sup.+
318.1917, found: 318.1926.
Synthesis 67
(3R,4R)-tert-butyl
4-(hydroxymethyl)-3-(methoxymethoxy)piperidine-1-carboxylate
(77)
##STR00181##
[0834] To a solution of (3R,4S)-1-tert-butyl 4-ethyl
3-(methoxymethoxy)piperidine-1,4-dicarboxylate (3.50 g, 11.0 mmol)
in CH.sub.2Cl.sub.2 (60 mL) at -20.degree. C. was added DIBAL-H (24
mL of a 1.0 M solution in hexane, 24.0 mmol). The solution was
stirred at -20.degree. C. for 1.5 h and then warmed to 22.degree.
C. The reaction mixture was quenched with Rochelle's salt and
stirred vigorously for 2 h. The organic layer was separated and the
aqueous phase was extracted with EtOAc (3.times.100 mL) and the
combined organic phases were dried over MgSO.sub.4 and concentrated
under reduced pressure to give a clear oil. Purification by column
chromatography (hexane:EtOAc 1:1) gave the title compound (2.35 g,
77%) as a clear oil.
[0835] R.sub.1=0.20 (hexane:EtOAc 1:1); [.alpha.].sub.D.sup.22+33.4
(c 1.11, CHCl.sub.3); .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
4.76 (d, J=6.7 Hz, 1H), 4.65 (d, J=6.7 Hz, 1H), 4.34 (brs, 1H),
4.04 (brs, 1H), 3.74-3.63 (m, 2H), 3.45-3.38 (m, 1H), 3.41 (s, 3H),
2.71-2.64 (m, 1H), 2.42 (brs, 2H) 1.71-1.64 (m, 2H), 1.45 (s, 9H),
1.39-1.34 (m, 1H); .sup.13C NMR (CDCl.sub.3, 100 MHz) .delta.
154.6, 96.1, 79.8, 75.5, 64.8, 55.9, 47.3, 44.2, 43.3, 28.4, 27.0;
HRMS (ESI) calc. for C.sub.13H.sub.25NO.sub.5 (M+Na).sup.+
298.1630, found: 298.1638.
Synthesis 68
(3R,4R)-tert-Butyl
4-(azidomethyl)-3-(methoxymethoxy)piperidine-1-carboxylate (37)
##STR00182##
[0837] To a solution of (3R,4R)-tert-butyl
4-(hydroxymethyl)-3-(methoxymethoxy)piperidine-1-carboxylate (2.25
g, 8.2 mmol) and DIPEA (7.15 mL, 41.0 mmol) in CH.sub.2Cl.sub.2 (80
mL) was added MsCl (1.91 mL, 24.6 mmol). After 2 h, the reaction
mixture was poured on water (50 mL) and extracted with EtOAc
(4.times.50 mL). The combined organic layers were washed NH.sub.4Cl
(50 mL) with brine (50 mL), dried over MgSO.sub.4, and concentrated
under reduced pressure to a light yellow oil. The residue was
dissolved in DMF (15 mL) and NaN.sub.3 (2.66 g, 41.0 mmol) and Nal
(122 mg, 0.82 mmol) were added and the mixture was heated
60.degree. C. After 48 h, the reaction mixture was poured on water
(50 mL) and extracted with EtOAc (3.times.50 mL). The combined
organic layers were washed water (20 mL) and brine (20 mL), dried
over MgSO.sub.4, and concentrated under reduced pressure to give a
light yellow oil. Purification by column chromatography
(hexane:EtOAc 2:1) gave 37 (1.92 g, 78%) as a clear oil. All
characterization data are identical to those reported in Synthesis
25.
[0838] Additional Compounds
[0839] The following additional compounds were prepared using
analogous methods.
TABLE-US-00004 Compound No. Structure Experimental Data PPDA-004
##STR00183## .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 7.88 (s,
1H), 5.56 (s, 1H), 3.76-3.66 (m, 2H), 3.54-3.39 (m, 4H), 3.11-2.95
(m, 2H), 2.82 (t, 1H, J = 11.3 Hz), 2.17 (ddd, J = 2.8, 6.1, 14.6
Hz, 1H), 1.85-1.67 (m, 8H), 1.36-1.30 (m, 10H), 1.11-1.03 (m, 2H
PPDA-005 ##STR00184## .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta.
7.81 (d, J = 8.7 Hz, 2H), 7.73 (s, 1H), 7.59 (d, J = 8.7 Hz, 2H),
5.52 (s, 1H), 3.97 (dd, J = 13.3, 3.3 Hz, 1H), 3.41 (td, J = 10.0,
4.3 Hz, 1H), 3.25- 3.20 (m, 2H), 3.15-3.11 (m, 1H), 3.07 (sept, J =
6.9 Hz, 1H), 2.74 (dd, J = 12.1, 3.2 Hz, 1H), 2.70 (s, 6H),
2.61-2.55 (m, 1H), 1.84-1.80 (m, 1H), 1.66- 1.49 (m, 2H), 1.30 (d,
J = 7.1 Hz, 3H), 1.28 (d, J = 7.1 Hz, 3H) HRMS (ESI) Calcd. for
C.sub.23H.sub.34N.sub.7O.sub.3S [M + H].sup.+, 488.2444, found
488.2459 PPDA-006 ##STR00185## .sup.1H NMR (CD.sub.3OD, 500 MHz)
.delta. 7.86 (dd, J = 8.6, 5.1 Hz, 2H), 7.61 (s, 1H), 7.36 (t, J =
8.7 Hz, 2H), 5.49 (s, 1H), 3.93 (dd, J = 14.5, 3.7 Hz, 1H), 3.81-
3.63 (m, 1H), 3.58-3.40 (m, 1H), 3.33-3.20 (m, 7H), 3.01 (sept, J =
6.9 Hz, 1H), 2.87 (td, J = 12.2, 3.4 Hz, 1H), 2.70 (dd, J = 12.0,
10.6 Hz, 1H), 2.67- 2.60 (m, 1H), 2.22-2.04 (m, 1H), 1.89 (dt, J =
10.4, 2.9 Hz, 1H), 1.75-1.61 (m, 4H), 1.34 (d, J = 6.8 Hz, 6H) HRMS
(ESI) Calcd. for C.sub.26H.sub.37N.sub.7O.sub.3FS [M + H].sup.+,
546.2663, found 546.2676 PPDA-008 ##STR00186## .sup.1H NMR
(CD.sub.3OD, 500 MHz) .delta. 8.00 (d, J = 7.6 Hz, 1H), 7.89 (d, J
= 8.0 Hz, 1H), 7.88 (s, 1H), 7.71 (s, 1H), 7.42 (t, J = 7.2 Hz,
1H), 7.38 (t, J = 6.8 Hz, 1H), 5.60 (s, 1H), 3.76- 3.70 (m, 1H),
3.65-3.6 (m, 1H), 3.45-3.42 (m, 1H), 3.37 (s, 2H), 3.11- 3.07 (m,
1H), 2.96-2.91 (m, 1H), 2.81-2.77 (m, 1H), 2.05-2.01 (m, 1H),
1.89-1.84 (m, 1H), 1.66- 1.62 (m, 1H), 1.31 (d, J = 6.8, 3H), 1.29
(d, J = 6.8, 3H) PPDA-011 ##STR00187## .sup.1H NMR (CD.sub.3OD, 500
MHz) .delta. 7.79 (s, 1H), 7.46- 7.44 (m, 1H), 7.42-7.40 (m, 1H),
7.31-7.29 (m, 2H), 5.57 (s, 1H) 4.73 (brs, 2H), 3.45 (s, 2H),
3.28-3.25 (m, 4H), 3.06 (sept, J = 6.8 Hz, 1H), 1.87-1.76 (m, 4H),
1.31 (d, J = 6.8 Hz, 6H) HRMS (ESI) Calcd. for
C.sub.22H.sub.30N.sub.6OCl [M + H].sup.+, 429.2170, found 429.216
PPDA-012 ##STR00188## .sup.1H NMR (CD.sub.3OD, 500 MHz) .delta.
7.90 (s, 1H), 7.64 (d, J = 10.0 Hz, 1H), 7.42 (d, J = 10 Hz, 1H),
7.36 (t, J = 10 Hz, 1H), 7.24 (t, J = 10 Hz, 1H), 5.45 (s, 1H),
4.81 (s, 2H), 3.49 (s, 2H), 3.34-3.28 (m, 4H), 3.15- 3.10 (m, 1H),
1.91-1.89 (m, 4H), 1.32 (6H, d, J = 6.8 Hz) .sup.13C NMR (125 MHz,
CD.sub.3OD) .delta. 155.5, 150.8, 143.9, 136.4, 134.8, 134.3,
132.4, 130.9, 130.1, 129.2, 124.1, 112.6, 68.7, 53.5, 47.2, 41.0,
32.1, 24.1, 23.5 HRMS (ESI) Calcd. for C.sub.22H.sub.30N.sub.6OBr
[M + H].sup.+, 473.1664, found 473.1667 PPDA-013 ##STR00189## HRMS
(ESI) Calcd. for C.sub.22H.sub.37N.sub.6O [M + H].sup.+, 401.3029,
found 401.3010 PPDA-014 ##STR00190## .sup.1H NMR (CD.sub.3OD, 500
MHz) .delta. 7.88 (s, 1H), 5.60 (s, 1H), 3.90-3.88 (m, 1H), 3.69
(d, J = 14.2 Hz, 1H), 3.63 (d, J = 14.2 Hz, 1H), 3.35-3.15 (m, 5H),
3.07 (sept, J = 6.8 Hz, 1H), 1.85-1.70 (m, 6H), 1.32 (d, J = 6.8
Hz, 6H), 1.30-1.25 (m, 4H), 1.11- 0.98 (m, 4H) PPDA-016
##STR00191## .sup.1H NMR (CD.sub.3OD, 500 MHz) .delta. 7.85 (m,
1H), 4.71 (m, 1H), 4.52 (s, 1H), 3.84- 3.80 (m, 1H), 3.51-3.38 (m,
4H), 3.07-3.02 (m, 1H), 1.82-1.64 (m, 6H), 1.35-1.30 (m, 6H), 1.23-
1.18 (m, 3H), 1.15-0.92 (m, 4H) .sup.13C-NMR (125 MHz, CD.sub.3OD)
.delta. 154.2, 151.3, 144.0, 134.8, 112.6, 69.7, 55.4, 53.4, 47.4,
39.0, 31.8, 27.4, 27.0, 24.2, 23.5 PPDA-017 ##STR00192## .sup.1H
NMR (CD.sub.3OD, 500 MHz) .delta. 7.93 (s, 1H), 7.73- 7.56 (m, 4H),
4.93 (s, 2H), 4.68 (s, 1H), 4.49 (s, 1H), 3.77-3.72 (m, 1H),
3.51-3.40 (m, 3H), 3.17- 3.12 (m, 2H), 1.32 (d, J = 6.8 Hz, 6H)
.sup.13C NMR (125 MHz, CD.sub.3OD) .delta. 154.4, 151.3, 144.2,
142.6, 135.0, 131.2, 130.9, 129.0, 126.9, 126.84, 126.81, 124.3,
112.7, 69.7, 55.3, 53.2, 47.2, 46.2, 30.8, 24.1, 23.5 PPDA-019
##STR00193## HRMS (ESI) Calcd. for C.sub.22H.sub.37N.sub.6O [M +
H].sup.+, 401.3029, found 401.3042 PPDA-020 ##STR00194## .sup.13C
NMR (125 MHz, CD.sub.3OD) .delta. 160.0, 155.8, 144.1, 143.1,
135.1, 126.6, 126.5, 112.9, 66.3, 65.4, 51.5, 46.2, 45.9, 31.7,
24.1, 23.5, 23.5; HRMS (ESI) Calcd. for C.sub.21H.sub.30N.sub.7O [M
+ H].sup.+, 396.2512, found 396.2527 PPDA-021 ##STR00195## .sup.1H
NMR (CD.sub.3OD, 500 MHz) .delta. 7.69 (s, 1H), 7.44- 7.42 (m, 1H),
7.40-7.38 (m, 1H), 7.29-7.26 (m, 2H), 5.15 (s, 1H), 4.64 (s, 2H),
3.55-3.34 (m, 6H), 3.16 (m, 2H), 3.02 (sept, J = 6.5 Hz, 1H),
1.96-1.85 (m, 2H), 1.30 (d, J = 6.5 Hz, 3H), 1.28 (d, J = 6.5 Hz,
3H) .sup.13C NMR (125 MHz, CD.sub.3OD) .delta. 159.5, 148.3, 146.0,
141.7, 136.0, 134.1, 130.7, 130.1, 129.3, 128.4, 113.7, 73.9, 64.8,
51.8, 50.8, 46.1, 44.8, 44.4, 31.5, 24.7, 23.9, 23.7 HRMS (ESI)
Calcd. for C.sub.22H.sub.30N.sub.6OCl [M + H].sup.+, 429.2170,
found 429.2175 PPDA-023 ##STR00196## .sup.1H NMR (CDCl.sub.3, 500
MHz) .delta. 7.95 (s, 1H), 7.47 (d, J = 7.5 Hz, 2H), 7.39 (t, J =
7.4 Hz, 2H), 7.33 (t, J = 7.1 Hz, 1H), 5.42 (s, 1H) 4.78 (s, 2H),
4.16 (s, 1H), 3.39-3.52 (m, 2H), 3.39 (s, 3H), 3.11-3.18 (m, 4H),
2.48 (d, J = 13.4 Hz, 1H), 1.85 (s, 1H), 1.35 (d, J = 6.0 Hz, 3H),
1.33 (d, J = 6.0 Hz, 3H) HRMS (ESI) Calcd. for
C.sub.22H.sub.30N.sub.6O [M + H].sup.+, 395.2559, found 395.2547
PPDA-024 ##STR00197## .sup.1H NMR (DMSO, 500 MHz) .delta. 9.55
(brs, 2H), 7.45-7.34 (m, 6H), 5.71 (brs, 1H), 4.66-4.62 (m, 2H),
4.12-4.09 (m, 1H), 3.20-3.11 (m, 9H), 2.48 (s, 1H), 2.15 (s, 1H),
1.62 (m, 1H), 1.20 (s, 6H); HRMS (ESI) Calcd. for
C.sub.22H.sub.31N.sub.6O.sub.2 [M + H].sup.+, 411.2508, found
411.2502 PPDA-025 ##STR00198## .sup.1H NMR (DMSO, 500 MHz) .delta.
9.55 (brs, 2H), 7.45-7.34 (m, 6H), 5.71 (brs, 1H), 4.66-4.62 (m,
2H), 4.12-4.09 (m, 1H), 3.20-3.11 (m, 9H), 2.48 (s, 1H), 2.15 (s,
1H), 1.62 (m, 1H), 1.20 (s, 6H) HRMS (ESI) Calcd. for
C.sub.22H.sub.31N.sub.6O.sub.2 [M + H].sup.+, 411.2508, found
411.2497 PPDA-027 ##STR00199## .sup.1H NMR (DMSO, 500 MHz) .delta.
9.86 (brs, 1H), 9.19 (brs, 1H), 8.30 (brs, 1H), 7.79 (s, 1H), 7.42
(d, J = 7.5 Hz, 2H), 7.35 (t, J = 7.2 Hz, 2H), 7.27 (t, J = 7.0 Hz,
1H), 5.37 (s, 1H), 4.60-4.53 (m, 3H), 3.83- 3.75 (m, 1H) 3.69 (dd,
J = 10.7, 7.4 Hz, 1H), 3.63 (dd, J = 10.7, 4.2 Hz, 1H), 3.55 (dd, J
= 11.8, 7.4 Hz, 1H), 3.36 (s, 3H), 3.14- 3.80 (m, 2H), 2.08-2.03
(m, 1H), 1.74-1.68 (m, 1H), 1.29 (d, J = 7.0 Hz, 6H) .sup.13C NMR
(125 MHz, DMSO) .delta. 143.0, 140.2, 139.4, 137.3, 127.9, 127.8,
126.6, 126.4, 126.3, 110.8, 70.2, 57.9, 57.0, 49.4, 48.2, 44.7,
39.0, 29.9, 22.5 HRMS (ESI) Calcd. for C.sub.22H.sub.30N.sub.6O [M
+ H].sup.+, 395.2559, found 395.2563 PPDA-028 ##STR00200## .sup.1H
NMR (DMSO, 500 MHz) .delta. 9.79 (brs, 1H), 9.15 (brs, 1H), 8.54
(brs, 1H), 7.83 (s, 1H), 7.43 (d, J = 6.5 Hz, 2H), 7.35 (t, J =
7.4, 7.0 Hz, 2H), 7.31- 7.25 (m, 1H), 5.43 (s, 1H), 4.65-4.60 (m,
1H), 4.64 (s, 2H), 4.59-4.56 (m, 1H), 3.76-3.66 (m, 3H), 3.57 (dd,
J = 11.6, 7.4 Hz, 1H), 3.13 (sept, J = 7 Hz, 1H), 3.08 (dd, J =
11.5, 6.0 Hz, 1H), 2.48-2.44 (m, 1H), 1.80-1.74 (m, 1H), 1.27 (d, J
= 7.0 Hz, 6H) HRMS (ESI) Calcd. for C.sub.21H.sub.28N.sub.6O [M +
H].sup.+, 381.2403, found 381.2410 PPDA-029 ##STR00201## .sup.1H
NMR (CD.sub.3OD, 500 MHz) .delta. 7.75 (s, 1H), 7.39- 7.32 (m, 5H),
5.32 (s, 1H), 4.58 (s, 2H), 4.14 (t, J =3.2 Hz, 1H), 4.02- 3.97 (m,
1H), 3.78 (dd, J = 15.4, 2.3 Hz, 1H), 3.52 (dd, J = 15.4, 6.4 Hz,
1H), 3.38 (m, 1H), 3.36 (s, 3H), 3.18 (dd, J = 12.4, 3.4 Hz, 1H),
3.04 (sept, J = 6.9 Hz, 1H), 2.29 (dd, J = 14.0, 6.8 Hz, 1H), 1.95
(ddd, J = 14.0, 10.4, 4.1 Hz, 1H), 1.34 (d, J = 6.9 Hz, 3H), 1.34
(d, J = 6.9 Hz, 3H) .sup.13C NMR (125 MHz, CD.sub.3OD) .delta.
158.3, 147.3, 143.7, 140.3, 137.5, 128.4, 127.1, 126.6, 112.2,
79.7, 72.4, 60.2, 55.3, 48.7, 45.1, 43.0, 32.1, 23.5, 22.5 HRMS
(ESI) Calcd. for C.sub.22H.sub.31N.sub.6O [M + H].sup.+, 395.2559,
found 395.2557 PPDA-030 ##STR00202## .sup.1H NMR (CD.sub.3OD, 500
MHz) .delta. 7.73 (s, 1H), 7.33- 7.27 (m, 5H), 5.38 (s, 1H), 4.55
(brs, 1H), 4.54 (s, 2H), 4.15-4.13 (m, 1H), 3.79 (br d, J = 14.9
Hz, 1H), 3.54 (dd, J = 15.1, 6.2 Hz, 1H), 3.21 (m, 2H), 3.03 (sept,
J = 6.9 Hz, 1H), 2.13 (dd, J = 13.4, 6.9 Hz, 1H), 2.01 (td, J =
6.9, 3.8 Hz, 1H), 1.31 (d, J = 6.9 Hz, 6H) HRMS (ESI) Calcd. for
C.sub.21H.sub.29N.sub.6O [M + H].sup.+, 381.2403, found 381.2410
PPDA-031 ##STR00203## .sup.1H NMR (CD.sub.3OD, 500 MHz) .delta.
7.96 (s, 1H), 7.42- 7.35 (m, 5H), 5.57 (s, 1H), 4.82 (s, 2H), 4.36-
4.32 (m, 1H), 3.79-3.77 (m, 3H), 3.67 (m, 1H), 3.50 (t, J = 7.4 Hz,
1H), 3.18 (sept, J = 6.8 Hz, 1H), 2.34 (m, 1H), 2.04 (m, 1H), 1.34
(d, J = 6.8 Hz, 6H) HRMS (ESI) Calcd. for C.sub.21H.sub.29N.sub.6O
[M + H].sup.+, 381.2403, found 381.2419 PPDA-032 ##STR00204##
.sup.1H NMR (CD.sub.3OD, 500 MHz) .delta. 7.75 (s, 1H), 7.44- 7.35
(m, 5H), 5.34 (s, 1H), 4.59 (s, 2H), 4.22 (m, 2H), 3.84 (dd, J =
14.7, 1.1 Hz, 1H), 3.68-3.63 (m, 2H), 3.29-3.25 (m, 2H), 3.02
(sept, J = 6.9 Hz, 1H), 1.33 (d, J = 6.9 Hz, 3H), 1.32 (d, J = 6.9
Hz, 3H) .sup.13C NMR (125 MHz, CD.sub.3OD) .delta. 158.4, 147.4,
143.5, 140.3, 137.5, 128.4, 127.2, 126.6, 112.2, 72.4, 72.1, 70.2,
48.6, 48.5, 45.0, 41.2, 23.4, 22.5; HRMS (ESI) Calcd. for
C.sub.21H.sub.29N.sub.6O.sub.2 [M + H].sup.+, 397.2352, found
397.2330
[0840] Biological Methods and Data
[0841] In vitro Kinase Assays and IC.sub.50 Determination
[0842] Purified recombinant CDK1/cycA1, CDK2/cycA1, CDK4/ cycD1,
CDK5/p35NCK, CDK6/cycD1, CDK7/CycH/MAT1, and CDK9/CycT1 were
purchased from ProQinase GmbH. Kinase assays were performed
according to manufacturer's protocols. Rb-CTF (ProQinase GmbH) (cat
number: 0040-0000-6) was used as the kinase substrate for CDK1,
CDK2, CDK4, and CDK6 kinases. RNA Polymerase II C-Terminal domain
(Pol II CTD) peptide (YSPTSPSYSPTSPSYSPTSPS) (Cambridge Research
Biochemicals) peptide was used for CDK7 and CDK9 kinase assays. A
luciferase assay (PKLight assay; Cambrex) was used to determine ATP
remaining at the end of the kinase reaction, which provides a
measure of kinase activity, according to the manufacturer's
protocols.
[0843] Kinase assays were carried out by incubation of increasing
amounts of test compound with purified recombinant CDK-Cyclin
complex, followed by measurement of free ATP remaining in the
reaction using the luciferase assay kit (PKLight, Cambrex) which
therefore provided a measure of inhibition for the specific
CDK.
[0844] 5 .mu.L of 1.times. kinase buffer (Cell Signalling
Technologies) was mixed with 200 ng CDK1, 200 ng CDK2, 50 ng CDK4,
100 ng CDK5, or 200 ng CDK6 with 5 .mu.g Rb-CTF and 300 ng CDK7 or
200 ng CDK9 with 500 .mu.M RNA Poll II CTD peptide. ATP at the
K.sub.m for each enzyme (0.16 .mu.M for CDK1; 0.58 .mu.M for CDK2;
18.7 .mu.M for CDK4; 1.8 .mu.M for CDK5; 20.9 .mu.M for CKD6; 4.1
.mu.M for CDK7; and 4 .mu.M for CDK9) was added to the reaction mix
and doubly distilled water was added to make the volume up to 39
.mu.L. The mixture was incubated at 30.degree. C. for 30 minutes.
The reaction is stopped with 20 .mu.L of Stop Solution (provided in
the PKLight kit) for 10 minutes at room temperature. Then 40 .mu.L
of the luciferase mix was added to the reaction mixture which was
incubated for a further 10 minutes at room temperature and measured
using the Tecan Infinite 2000 plate reader. GraphPad Prism Software
was used to generate standard curve and determine the IC.sub.50's
for each CDK.
[0845] CDK activities for PPDA-001 were determined using an in
vitro kinase assay, as described above. The IC.sub.50 values
(.mu.mol/L) are shown in the following table. The results of three
experiments are reported, along with standard errors of the means
(SEM) (.mu.mol/L).
TABLE-US-00005 TABLE 1 IC.sub.50 Data for PPDA-001 Kinase IC.sub.50
(.mu.mol/L) SEM (.mu.mol/L) CDK1 1.52 0.04 CDK2 0.58 0.1 CDK4 42.1
0.9 CDK5 9.0 0.11 CDK6 32.1 0.8 CDK7 0.041 0.04 CDK9 1.1 0.03
[0846] In vitro kinase inhibition data (CDK1, CDK2, CDK7) and
selectivity (CDK1/7, CDK2/7) data are summarised in the following
table.
TABLE-US-00006 TABLE 2 IC.sub.50 Data for Various Compounds In
vitro Kinase Inhibition Selectivity CDK1 CDK2 CDK7 CDK1/ CDK2/
IC.sub.50 IC.sub.50 IC.sub.50 CDK7 CDK7 Compound (nM) (nM) (nM)
(fold) (fold) PPDA-001 1520 580 41 37 14 PPDA-002 1380 2030 18 77
113 PPDA-003 1910 114 47 41 2.5 PPDA-004 1820 1290 940 1.9 1.4
PPDA-005 213 38 111 1.9 0.3 PPDA-006 405 178 40 10 4.5 PPDA-007
1146 503 461 2.5 1.1 PPDA-008 2950 42 120 25 0.4 PPDA-009 559 459
462 1.2 1.0 PPDA-010 3625 115 788 4.6 0.1 PPDA-011 143 98 414 0.3
0.2 PPDA-012 74 118 484 0.2 0.2 PPDA-013 330 344 80 4.1 4.3
PPDA-014 8680 481 975 8.9 0.5 PPDA-015 3950 1290 27 146 48 PPDA-016
1460 4550 246 5.9 18 PPDA-017 1350 754 1890 0.7 0.4 PPDA-018 41 1
14 2.9 0.1 PPDA-019 80 9 29 2.8 0.3 PPDA-020 441 22 44 10 0.5
PPDA-021 177 22 81 2.2 0.3 PPDA-022 1900 568 73 26 7.8 PPDA-023
1200 450 60 20 7.5 PPDA-024 844 1027 150 5.6 6.8 PPDA-025 285 106
77 3.7 1.4 PPDA-026 162 203 89 1.8 2.3 PPDA-027 253 680 18 14 38
PPDA-028 4010 262 43 93 6.1 PPDA-029 -- 2840 1007 -- 2.8 PPDA-030
-- 999 59 -- 17 PPDA-031 -- 393 473 -- 0.8 PPDA-032 -- 1500 305 --
4.9
[0847] Cell Growth Inhibitions Assays
[0848] All cells were purchased from the American Type Culture
Collection (ATCC) and MCF7 cells were routinely cultured in
Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10%
fetal calf serum (FCS) (First Link) and HCT116 cells were routinely
cultured in Roswell Park Memorial Insititute Medium (RPMI)
supplemented with 10% FCS. Cell growth was assessed using a known
sulforhodamine B assay (see, e.g., Skehan et al., 1990).
[0849] MCF-7 cells purchased from ATCC (USA) were routinely
passaged in DMEM, supplemented with 10% FCS and HCT116 cells
purchased from ATCC (USA) were routinely passaged in RPMI
supplemented with 10% FCS and kept in a 37.degree. C. incubator
with 5% CO2. The growth assay for both cell lines was performed in
the appropriate media using using the exact same protocol as
described here. For the growth assay, 5000 cells were seeded into
each well of 96-well plates in DMEM containing 10% FCS. Test
compounds prepared in DMSO were added to the medium at
concentrations ranging from 0.00038-100 .mu.M. The cells were
incubated for a further 72 hours, at which time they were fixed by
the addition of 100 .mu.L/well of ice-cold 40% trichloroacetic acid
(TCA). The plates were left for 1 hour at 4.degree. C., washed in
water and 100 .mu.L of 0.4% (w/v) sulforhodamine (SRB;
Sigma-Aldrich, UK) prepared in 1% acetic acid was added. Plates
were washed in 1% acetic acid to remove excess SRB reagent, air
dried, and bound dye was solubilized by the addition of 100 .mu.L
of 10 mM tris base. The plates were read at 492 nm using a plate
reader. The optical densities (OD) at 492 nm were plotted to
determine the concentration of test compound at which 50%
inhibition of growth is observed (using GraphPad Prism).
[0850] PPDA-001 inhibited the growth of breast (MCF7) and
colorectal (HCT116) cancer cell lines with Gl.sub.50 values of
<1 .mu.mol/L.
[0851] The data are summarised in the following table.
TABLE-US-00007 TABLE 3 Growth Inhibition, GI.sub.50 (.mu.M)
Compound No MCF7 HCT116 PPDA-001 0.96 0.63 PPDA-002 1.5 5.1
PPDA-003 -- -- PPDA-004 2.7 2.5 PPDA-005 4.22 7.2 PPDA-006 8.4 10.0
PPDA-007 2.1 4.1 PPDA-008 3.1 5.1 PPDA-009 38.9 86.6 PPDA-010 11.9
25.0 PPDA-011 0.3 1.6 PPDA-012 0.5 1.6 PPDA-013 1.2 1.1 PPDA-014
45.6 53.5 PPDA-015 4.1 12.7 PPDA-016 13.1 7.4 PPDA-017 12.8 11.4
PPDA-018 0.1 0.9 PPDA-019 0.3 0.4 PPDA-020 0.6 1.5 PPDA-021 0.2 1.2
PPDA-022 3.7 10.4 PPDA-023 3.7 5.6 PPDA-024 12.2 46.4 PPDA-025 10.1
23.7 PPDA-026 2.0 10.7 PPDA-027 3.4 8.9 PPDA-028 3.3 7.3 PPDA-029
17.6 >100 PPDA-030 20.2 >100 PPDA-031 5.6 20.4 PPDA-032 --
--
[0852] NCI Screening
[0853] To extend the analysis to a more extensive set of cancer
cell lines, PPDA-001 was submitted to the National Cancer
Institute's Division of Cancer Treatment and Diagnosis in vitro
screen of human tumor cell lines
(http://dtp.nci.nih.gov/branches/btb/ivclsp.html).
[0854] The results are shown graphically in FIG. 1.
[0855] FIG. 1 is a graph of percentage growth inhibition as a
function of the base-10 logarithm of the molar concentration of the
test compound, PPDA-001, as determined by the NCI60 cancer cell
line screen. Each line represents one cell line.
[0856] The screen demonstrated that PPDA-001 caused inhibition of
all 60 cancer cell lines (mean Gl.sub.50=0.28 .mu.mol/L; range
Gl.sub.50=0.04 to 2.1 .mu.mol/L).
[0857] HCT116 Tumour Xenograft Study
[0858] Animals (female Balb/c nu/nu mice) were randomized to 4 arms
(15 animals in each arm), with tumor-bearing animals being treated
PO by oral gavage, with PPDA-001 using 5% DMSO in PBS as vehicle.
Once tumors reached a volume of 100-200 mm.sup.3, the animals were
either left untreated, or treated with vehicle, 50 mg/kg PPDA-001
bi-daily (bd) (8-hours between 1.sup.st and 2.sup.nd administration
daily), or treated once daily with 100 mg/kg PPDA-001.
[0859] The results are illustrated in FIG. 2.
[0860] FIG. 2 is a graph of relative tumour volume as a function of
time in the HCT116 tumour xenograft study, for vehicle control
(squares), 50 mg/kg/bi-daily (triangles), and 100 mg/kg once daily
(crosses). Error bars represent standard errors of the mean
(SEM).
[0861] Animal weights in the PPDA-001 treatment arms fell over the
course of the study, reaching 92%, compared to vehicle treated
animals whose weight fell to 98%.
[0862] The results are illustrated in FIG. 3.
[0863] FIG. 3 is a graph of percent body weight as a function of
time in the HCT116 tumour xenograft study, for vehicle control
(squares), 50 mg/kg/bi-daily (triangles), and 100 mg/kg once daily
(crosses).
[0864] PPDA-001 substantially reduced tumor growth at the 50
mg/kg/bi-daily and 100 mg/kg once daily treatment regimens,
demonstrating a 65% reduction in tumor growth in the PPDA-001
treatment arms, as compared with the control arms. There was no
difference in tumor growth between these two doses
(p<0.001).
[0865] Comparison Studies
[0866] The following compounds were prepared for comparison
purposes, in particular, for comparison with PPDA-001. These
compounds differ from PPDA-001 invention by (a) the absence of an
"oxy" substituent, in the case of XX-01, or (b) the absence of a
nitrogen-containing heterocyclic group, in the case of XX-02.
##STR00205##
[0867] Corresponding data for PPDA-001 and these comparison
compounds are summarised in the following tables.
TABLE-US-00008 TABLE 5 IC.sub.50 Data for Various Compounds In
vitro Kinase Inhibition Selectivity CDK1 CDK2 CDK7 CDK1/ CDK2/
IC.sub.50 IC.sub.50 IC.sub.50 CDK7 CDK7 Compound (nM) (nM) (nM)
(fold) (fold) PPDA-001 1520 580 41 37 14 XX-01 1660 1520 310 5.4
4.9 XX-02 -- 1695 687 -- 2.5
[0868] As can be seen from the data, PPDA-001 has a substantially
greater selectivity for CKD7, as compared to both CKD1 and CKD2,
than the structurally similar comparison compounds.
[0869] CDK7 selectively is desirable since CDK7 is a member of a
large family of protein kinases with important developmental and
cellular roles, comprising 25 members in man. CDK7 regulates cell
cycle progression by phosphorylating and thereby activating cell
cycle CDKs. In addition, CDK7 phosphorylates RNA polymerase II to
facilitate gene transcription. Although CDK7 is an essential gene
during development, in the adult CDK7 is not essential, as
demonstrated in knockout mice, where its deletion demonstrates no
phenotype in tissues with low proliferative index. However,
re-population of cells in tissues with high cellular turnover is
associated with adult stem cell depletion and premature ageing
(see, e.g., Ganuza et al., 2012). Hence, treatment with CDK7
inhibitors is expected to be associated with fewer side
effects.
TABLE-US-00009 TABLE 6 Growth Inhibition, GI.sub.50 (.mu.M)
Compound No MCF7 HCT116 PPDA-001 0.96 0.63 XX-01 12.2 18.7 XX-02 47
>100
[0870] As can be seen from the data, PPDA-001 has a substantially
greater growth inhibition than the structurally similar comparison
compounds.
[0871] The foregoing has described the principles, preferred
embodiments, and modes of operation of the present invention.
However, the invention should not be construed as limited to the
particular embodiments discussed. Instead, the above-described
embodiments should be regarded as illustrative rather than
restrictive. It should be appreciated that variations may be made
in those embodiments by workers skilled in the art without
departing from the scope of the present invention.
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